Lxr modulators

ABSTRACT

Compounds, pharmaceutically acceptable salts, isomers, or prodrugs thereof, of the invention are disclosed, which are useful as modulators of the activity of liver X receptors (LXR). Pharmaceutical compositions containing the compounds and methods of using the compounds are also disclosed.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a Continuation of U.S. patent application Ser. No.14/703,247, filed May 4, 2015, which is a Continuation of U.S. patentapplication Ser. No. 14/085,926, filed Nov. 21, 2013, which is aContinuation of U.S. patent application Ser. No. 13/319,937 filed Nov.30, 2011, now U.S. Pat. No. 8,618,154, which is a 35 U.S.C. §371National Phase Application of International Application Serial No.PCT/US2010/036211, filed May 26, 2010, which claims priority to U.S.Provisional Patent Application No. 61/181,736, filed May 28, 2009. Thedisclosures of which are hereby incorporated in their entirety byreference.

BACKGROUND OF THE INVENTION Field of the Invention

This invention relates to compounds that modulate the activity of liverX receptors (LXRs). The invention also provides pharmaceuticalcompositions comprising the compounds of the invention and methods ofutilizing those compositions for modulating the activity of liver Xreceptor. In particular, imidazole isomers and derivatives are providedfor modulating the activity of LXRs.

Nuclear Receptors

Nuclear receptors are a superfamily of regulatory proteins that arestructurally and functionally related and are receptors for, e.g.,steroids, retinoids, vitamin D and thyroid hormones (see, e.g., Evans(1988) Science 240:889-895). These proteins bind to cis-acting elementsin the promoters of their target genes and modulate gene expression inresponse to ligands for the receptors.

Nuclear receptors can be classified based on their DNA bindingproperties (see, e.g., Evans, supra and Glass (1994) Endocr. Rev.15:391-407). For example, one class of nuclear receptors includes theglucocorticoid, estrogen, androgen, progestin and mineralocorticoidreceptors which bind as homodimers to hormone response elements (HREs)organized as inverted repeats (see, e.g., Glass, supra). A second classof receptors, including those activated by retinoic acid, thyroidhormone, vitamin D₃, fatty acids/peroxisome proliferators (i.e.,peroxisome proliferator activated receptors or PPARs) and ecdysone, bindto HREs as heterodimers with a common partner, the retinoid X receptors(i.e., RXRs, also known as the 9-cis retinoic acid receptors; see, e.g.,Levin et al. (1992) Nature 355:359-361 and Heyman et al. (1992) Cell68:397-406).

RXRs are unique among the nuclear receptors in that they bind DNA as ahomodimer and are required as a heterodimeric partner for a number ofadditional nuclear receptors to bind DNA (see, e.g., Mangelsdorf et al.(1995) Cell 83:841-850). The latter receptors, termed the class IInuclear receptor subfamily, include many which are established orimplicated as important regulators of gene expression.

There are three RXR genes (see, e.g., Mangelsdorf et al. (1992) GenesDev. 6:329-344), coding for RXRα, β, and γ, all of which are able toheterodimerize with any of the class II receptors, although there appearto be preferences for distinct RXR subtypes by partner receptors in vivo(see, e.g., Chiba et al. (1997) Mol. Cell. Biol. 17:3013-3020). In theadult liver, RXRa is the most abundant of the three RXRs (see, e.g.,Mangelsdorf et al. (1992) Genes Dev. 6:329-344), suggesting that itmight have a prominent role in hepatic functions that involve regulationby class II nuclear receptors. See also, Wan et al. (2000) Mol. Cell.Biol. 20:4436-4444.

LXR_(α) and LXR_(β)

LXR_(α) is found predominantly in the liver, with lower levels found inkidney, intestine, spleen and adrenal tissue (see, e.g., Willy, et al.(1995) Gene Dev. 9(9):1033-1045). LXR_(β) is ubiquitous in mammals andwas found in nearly all tissues examined. LXRs are activated by certainnaturally occurring, oxidized derivatives of cholesterol (see, e.g.,Lehmann, et al. (1997) J. Biol. Chem. 272(6):3137-3140). LXR_(α) isactivated by oxycholesterol and promotes cholesterol metabolism (Peet etal. (1998) Cell 93:693-704). Thus, LXRs appear to play a role in, e.g.,cholesterol metabolism (see, e.g., Janowski, et al. (1996) Nature383:728-731).

The nuclear receptor LXR plays a critical role in coordinate control ofbile acid, cholesterol, and triglyceride metabolism to maintain lipidhomeostasis. LXRs and bile acid/oxysterol-regulated genes are potentialtargets for developing drug therapies for lowering serum cholesterol andtreating cardiovascular and liver diseases. Compounds with activity atLXR can have profound effects on lipid homeostasis, and can moreeffectively control disease or disorders in which LXR is implicated.This is accomplished through regulation of multiple genes involved incholesterol homeostasis including Cyp7a1, a member of the cytochromep450 family of enzymes and the rate limiting step in bile acidsynthesis, as well as the ABC membrane transporters ABCA1, ABCG1, ABCG5,and ABCG8. ABCA1 is critical in the efflux of cholesterol andphospholipids to lipid-poor lipoproteins such as ApoA-I thuscontributing to an increase in plasma HDL levels. In addition, ABCG5 andABCG8 appear to mediate decreased intestinal absorption of cholesteroland facilitate cholesterol efflux from liver cells into the bile.Unfortunately, in addition to the anti-atherogenic effect of LXRagonists, studies in cell culture and animal model systems havedemonstrated that LXR agonists increase plasma triglyceride levels andhepatic lipogenesis and promote the increased production of VLDLlipoprotein particles. Schultz et al., Genes & Development 14:2831-2838(2000); Repa et al. Genes & Development 14:28119-2830 (2000). Strategiesto minimize the undesirable lipid effects include identifying LXRβselective compounds that are also partial agonists. Partial agonists candisplay tissue-specific activation or repression of nuclear receptors,as was demonstrated for the anti-estrogen tamoxifen, which functions asan antagonist of estrogen signaling in breast tissue and an agonist inthe uterus. Characterization of LXR isoform-specific null mice indicatethat LXRα is the predominant mediator of LXR activity in the liver. Inmacrophages, however, LXRβ alone is sufficient to mediate the effects ofLXR ligands on target gene expression. Therefore compounds with limitedLXRα activity should have anti-atherogenic activity while limitingunwanted hepatic effects.

SUMMARY OF THE INVENTION

Thus, we recognized that there is a need for compounds, compositions andmethods of modulating the activity of the LXR nuclear receptors in waysthat separate the desirable effects on cholesterol metabolism andatherogenesis from increased plasma triglyceride levels and an increasein hepatic lipogenesis. Although full agonists of LXR cause both thedesirable and undesirable effects, the present invention describescompounds that have a beneficial separation between the two, and thushave an improved therapeutic index between increased reverse cholesteroltransport and detrimental effects on plasma triglycerides andLDL-cholesterol.

In one aspect, the present invention comprises compounds or anindividual isomer or mixture of isomers, an isotope or apharmaceutically acceptable salt thereof, which are useful as modulatorsof the activity of liver X receptors (LXRs).

Compounds for use in compositions and methods for modulating theactivity of nuclear receptors are provided. In particular, compounds ofthe invention which are useful for modulating the liver X receptors,LXR_(α) and LXR_(β), and in particular, LXR_(β).

In one aspect, the compounds provided herein are agonists of LXR. Inanother aspect, the compounds provided herein are antagonists of LXR.Agonists that exhibit low efficacy are, in certain aspects, antagonists.

Another aspect of this invention is directed to methods of treating,inhibiting, or ameliorating the symptoms of a disease or disorder thatis modulated or otherwise affected by LXR activity or in which LXRactivity is implicated, comprising administering to a subject in needthereof a therapeutically effective amount of a compound of the presentinvention or an individual isomer or mixture of isomers or apharmaceutically acceptable salt thereof.

Another aspect of this invention is directed to methods of modulatingcholesterol metabolism to a subject in need thereof, comprisingadministering an effective cholesterol metabolism-modulating amount of acompound of the present invention or an individual isomer or mixture ofisomers or a pharmaceutically acceptable salt thereof.

Another aspect of this invention is directed to methods of preventing ortreating atherosclerosis in a subject in need thereof, comprisingadministering an effective cholesterol level-reducing amount of acompound of the present invention or an individual isomer or mixture ofisomers or a pharmaceutically acceptable salt thereof.

Another aspect of this invention is directed to methods of modulatingLXR activity to a subject in need thereof, comprising contacting thenuclear receptor with a compound of the present invention or anindividual isomer or mixture of isomers or a pharmaceutically acceptablesalt thereof.

Another aspect of this invention is directed to methods of treating,inhibiting or ameliorating one or more symptoms of hypocholesterolemiain a subject in need thereof, comprising administering a therapeuticallyeffective amount of a compound of the present invention or an individualisomer or mixture of isomers or a pharmaceutically acceptable saltthereof.

Another aspect of this invention is directed to methods of increasingcholesterol efflux from cells of a subject in need thereof, comprisingadministering an effective cholesterol efflux-increasing amount of acompound of the present invention or an individual isomer or mixture ofisomers or a pharmaceutically acceptable salt thereof.

Another aspect of this invention is directed to methods of increasingthe expression of ATP-Binding Cassette A1 (ABCA1) and ATP-BindingCassette G1 (ABCG1) in the cells of a subject in need thereof,comprising administering an effective ABCA1 and ABCG1expression-increasing amount of a compound of the present invention oran individual isomer or mixture of isomers or a pharmaceuticallyacceptable salt thereof.

Another aspect of this invention is directed to methods of treating,inhibiting, or ameliorating one or more symptoms of a disease ordisorder which is affected by cholesterol or bile acid levels,comprising administering to a subject in need thereof a therapeuticallyeffective amount of a compound of the present invention or an individualisomer or mixture of isomers or a pharmaceutically acceptable saltthereof.

Another aspect of this invention is directed to pharmaceuticalcompositions comprising a compound of the present invention or anindividual isomer or mixture of isomers or a pharmaceutically acceptablesalt thereof and at least one pharmaceutically acceptable carrier orexcipient.

Another aspect of this invention is directed to regulation of reversecholesterol transport and inflammatory signaling pathways that areimplicated in human disease pathology including atherosclerosis andassociated diseases such as myocardial infarction and ischemic stroke ina subject in need thereof, comprising administering an effective reversecholesterol transport and inflammatory signaling pathways regulatingamount of a compound of the present invention or an individual isomer ormixture of isomers or a pharmaceutically acceptable salt thereof.

Another aspect of this invention is directed to treatment of themetabolic syndrome which comprises a constellation of disorders of thebody's metabolism including obesity, hypertension, insulin resistance,and diabetes including treatment of diseases resulting from compromisedmetabolism and immunity including atherosclerosis and diabetes as wellas autoimmune disorders and diseases in a subject in need thereof,comprising administering a therapeutically effective amount of acompound of the present invention or an individual isomer or mixture ofisomers or a pharmaceutically acceptable salt thereof.

Another aspect of this invention is directed to treatment of theatherosclerosis, insulin resistance, osteoarthritis, stroke,hyperglycemia, dyslipidemia, psoriasis, age and UV exposure-dependentskin wrinkling, diabetes, cancer, Alzheimer's disease, inflammation,immunological disorders, lipid disorders, obesity, macular degeneration,conditions characterized by a perturbed epidermal barrier function,conditions of disturbed differentiation or excess proliferation of theepidermis or mucous membrane, or cardiovascular disorders in a subjectin need thereof, comprising administering a therapeutically effectiveamount of a compound of the present invention or an individual isomer ormixture of isomers or a pharmaceutically acceptable salt thereof.

DETAILED DESCRIPTION OF THE INVENTION

In one aspect, the present invention comprises a compound of formula I,

-   -   or a pharmaceutically acceptable salt thereof, wherein:    -   R¹ is chloro, fluoro, methyl or trifluoromethyl;    -   R² is H or methyl;    -   R³ is H or methyl;    -   R⁴ is H, chloro, fluoro, or methyl; and    -   n is 1, or 2.

In some embodiments, the compound of formula 1 is one in which R² and R³are methyl.

In some embodiments, the compound of formula I is one in which R¹ ischloro or fluoro. In some such embodiments, R² and R³ are methyl.

In some embodiments, the compound of formula I is one in which R⁴ isfluoro. In some such embodiments, R¹ is chloro or fluoro and R² and R³are methyl

In some embodiments, the compound of formula I is one in which R² and R³are H. In some such embodiments, R¹ is chloro or fluoro.

In some embodiments, the compound of formula I is one in which R² ismethyl and R³ is H. In some such embodiments, n is 2, R¹ is chloro, andR⁴ is fluoro.

In another aspect, the invention comprises a compound of structuralformula 1 according to any of the foregoing embodiments together withone or more pharmaceutically acceptable carriers, excipients, ordiluents.

In another aspect, the invention comprises a method of treating adisease or disorder comprising administering to a subject in needthereof a therapeutically effective amount of (a) a compound ofstructural formula I according to any of the foregoing embodiments or(b) a pharmaceutical composition comprising a compound of structuralformula I according to any of the foregoing embodiments together withone or more pharmaceutically acceptable carriers, excipients, ordiluents, wherein the disease or disorder is atherosclerosis,hypercholesterolemia, hyperlipoproteinemia, hypertriglyceridemia,lipodystrophy, hyperglycemia, diabetes mellitus, dyslipidemia,atherosclerosis, gallstone disease, acne vulgaris, acneiform skinconditions, diabetes, Parkinson's disease, cancer, Alzheimer's disease,inflammation, immunological disorders, lipid disorders, obesity,conditions characterized by a perturbed epidermal barrier function,conditions of disturbed differentiation or excess proliferation of theepidermis or mucous membrane, or cardiovascular disorders.

In some embodiments, the disease or disorder for treatment ishypercholesterolemia, hyperlipoproteinemia, hypertriglyceridemia,lipodystrophy, hyperglycemia, atherosclerosis, diabetes mellitus, ordyslipidemia.

In another embodiment, the present invention comprises a compound,isotope, or pharmaceutically acceptable salt thereof, selected from:

No. Name 12-(1-(3′-fluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-2-(2-(2-fluorophenyl)propan-2-yl)-1H-imidazol-4-yl)propan-2-ol; 22-(2-(2-(2-chloro-6-fluorophenyl)propan-2-yl)-1-(3′-fluoro-4′-(hydroxymethyl)-3-methyl-5′-(methylsulfonyl)biphenyl-4-yl)-1H-imidazol-4-yl)propan-2-ol; 32-(1-(3-chloro-3′-fluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-2-(2-(2,6-dichlorophenyl)propan-2-yl)-1H-imidazol-4-yl)propan-2-ol; 42-(2-(2-(2-chloro-3-fluorophenyl)propan-2-yl)-1-(3′-fluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-1H-imidazol-4-yl)propan-2-ol; 52-(2-(2-(2,6-dichlorophenyl)propan-2-yl)-1-(3′-fluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-1H-imidazol-4-yl)propan-2-ol; 62-(2-(2-(2-Chloro-phenyl)propan-2-yl)-1-(3,3′-difluoro-4′-hydroxymethyl-5′-(methylsulfonyl)biphenyl-4-yl)-1H-imidazol-4-yl)-propan-2-ol; 72-(2-(2-(2-chloro-6-fluorophenyl)propan-2-yl)-1-(3,3′-difluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-1H-imidazol-4-yl)propan-2-ol; 82-{1-(3,3′-Difluoro-4′-hydroxymethyl-5′-methanesulfonyl-biphenyl-4-yl)-2-[2-(2-fluorophenyl)propan-2-yl]-1H-imidazol-4-yl}-propan-2-ol; 92-(2-(2-(2,6-dichlorophenyl)propan-2-yl)-1-(3,3′-difluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-1H-imidazol-4-yl)propan-2-ol; 102-(2-(2-(2,6-dichlorophenyl)propan-2-yl)-1-(3,3′-difluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-1H-imidazol-4-yl)[(¹³CD₃)₂]propan-2-ol;11 2-(2-(2,4-dichlorobenzyl)-1-(3,3′-difluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-1H-imidazol-4-yl)propan-2-ol; 122-(1-(3,3′-difluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-2-(2-(trifluoromethyl)benzyl)-1H-imidazol-4-yl)propan-2-ol; 132-(1-(3-chloro-3′-fluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-2-(2-chloro-4-fluorobenzyl)-1H-imidazol-4-yl)propan-2-ol; 142-(2-(2-chloro-4-fluorobenzyl)-1-(3,3′-difluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-1H-imidazol-4-yl)propan-2-ol; 152-(2-(2,4-dichlorobenzyl)-1-(3′-fluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-1H-imidazol-4-yl)propan-2-ol; 162-(1-(3,3′-difluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-2-(2-fluorobenzyl)-1H-imidazol-4-yl)propan-2-ol; 172-(1-(3′-fluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-2-(2-methylbenzyl)-1H-imidazol-4-yl)propan-2-ol; 182-(2-(2,6-dichlorobenzyl)-1-(3′-fluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-1H-imidazol-4-yl)propan-2-ol; 192-[2-(2-Chloro-5-fluoro-benzyl)-1-(3′-fluoro-4′-hydroxymethyl-5′-methanesulfonyl-biphenyl-4-yl)-1H-imidazol-4-yl]-propan-2-ol; 202-[2-(2-Chloro-benzyl)-1-(3,3′-difluoro-4′-hydroxymethyl-5′-methanesulfonyl-biphenyl-4-yl)-1H-imidazol-4-yl]-propan-2-ol; or 212-{2-[1-(2,6-dichlorophenyl)ethyl]-1-[3,3′-difluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl]-1H-imidazol-4-yl}propan-2-ol.

In another embodiment, the present invention comprises a compound,isotope, or pharmaceutically acceptable salt thereof, selected from:

No. Name 12-(1-(3′-fluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-2-(2-(2-fluorophenyl)propan-2-yl)-1H-imidazol-4-yl)propan-2-ol; 22-(2-(2-(2-chloro-6-fluorophenyl)propan-2-yl)-1-(3′-fluoro-4′-(hydroxymethyl)-3-methyl-5′-(methylsulfonyl)biphenyl-4-yl)-1H-imidazol-4-yl)propan-2-ol; 32-(1-(3-chloro-3′-fluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-2-(2-(2,6-dichlorophenyl)propan-2-yl)-1H-imidazol-4-yl)propan-2-ol; 42-(2-(2-(2-chloro-3-fluorophenyl)propan-2-yl)-1-(3′-fluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-1H-imidazol-4-yl)propan-2-ol; 52-(2-(2-(2,6-dichlorophenyl)propan-2-yl)-1-(3′-fluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-1H-imidazol-4-yl)propan-2-ol; 62-(2-(2-(2-Chloro-phenyl)propan-2-yl)-1-(3,3′-difluoro-4′-hydroxymethyl-5′-(methylsulfonyl)biphenyl-4-yl)-1H-imidazol-4-yl)-propan-2-ol; 72-(2-(2-(2-chloro-6-fluorophenyl)propan-2-yl)-1-(3,3′-difluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-1H-imidazol-4-yl)propan-2-ol; 82-{1-(3,3′-Difluoro-4′-hydroxymethyl-5′-methanesulfonyl-biphenyl-4-yl)-2-[2-(2-fluorophenyl)propan-2-yl]-1H-imidazol-4-yl}-propan-2-ol; or 92-(2-(2-(2,6-dichlorophenyl)propan-2-yl)-1-(3,3′-difluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-1H-imidazol-4-yl)propan-2-ol; 212-{2-[1-(2,6-dichlorophenyl)ethyl]-1-[3,3′-difluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl]-1H-imidazol-4-yl}propan-2-ol.

In another embodiment, the present invention comprises a compound,isotope, or pharmaceutically acceptable salt thereof, selected from:

No. Name 112-(2-(2,4-dichlorobenzyl)-1-(3,3′-difluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-1H-imidazol-4-yl)propan-2-ol; 122-(1-(3,3′-difluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-2-(2-(trifluoromethyl)benzyl)-1H-imidazol-4-yl)propan-2-ol; 132-(1-(3-chloro-3′-fluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-2-(2-chloro-4-fluorobenzyl)-1H-imidazol-4-yl)propan-2-ol; 142-(2-(2-chloro-4-fluorobenzyl)-1-(3,3′-difluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-1H-imidazol-4-yl)propan-2-ol; 152-(2-(2,4-dichlorobenzyl)-1-(3′-fluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-1H-imidazol-4-yl)propan-2-ol; 162-(1-(3,3′-difluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-2-(2-fluorobenzyl)-1H-imidazol-4-yl)propan-2-ol; 172-(1-(3′-fluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-2-(2-methylbenzyl)-1H-imidazol-4-yl)propan-2-ol; 182-(2-(2,6-dichlorobenzyl)-1-(3′-fluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-1H-imidazol-4-yl)propan-2-ol; 192-[2-(2-Chloro-5-fluoro-benzyl)-1-(3′-fluoro-4′-hydroxymethyl-5′-methanesulfonyl-biphenyl-4-yl)-1H-imidazol-4-yl]-propan-2-ol; or 202-[2-(2-Chloro-benzyl)-1-(3,3′-difluoro-4′-hydroxymethyl-5′-methanesulfonyl-biphenyl-4-yl)-1H-imidazol-4-yl]-propan-2-ol.

In another embodiment, the present invention comprises a compound,isotope, or pharmaceutically acceptable salt thereof, selected from:

No. Name 32-(1-(3-chloro-3′-fluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-2-(2-(2,6-dichlorophenyl)propan-2-yl)-1H-imidazol-4-yl)propan-2-ol; 42-(2-(2-(2-chloro-3-fluorophenyl)propan-2-yl)-1-(3′-fluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-1H-imidazol-4-yl)propan-2-ol; 52-(2-(2-(2,6-dichlorophenyl)propan-2-yl)-1-(3′-fluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-1H-imidazol-4-yl)propan-2-ol; 92-(2-(2-(2,6-dichlorophenyl)propan-2-yl)-1-(3,3′-difluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-1H-imidazol-4-yl)propan-2-ol; or 212-{2-[1-(2,6-dichlorophenyl)ethyl]-1-[3,3′-difluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl]-1H-imidazol-4-yl}propan-2-ol.

In another embodiment, the present invention comprises a compound,isotope, or pharmaceutically acceptable salt thereof, which is2-(1-(3-chloro-3′-fluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-2-(2-(2,6-dichlorophenyl)propan-2-yl)-1H-imidazol-4-yl)propan-2-ol.

In another embodiment, the present invention comprises a compound,isotope, or pharmaceutically acceptable salt thereof, which is2-(2-(2-(2-chloro-3-fluorophenyl)propan-2-yl)-1-(3′-fluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-1H-imidazol-4-yl)propan-2-ol.

In another embodiment, the present invention comprises a compound,isotope, or pharmaceutically acceptable salt thereof, which is2-(2-(2-(2,6-dichlorophenyl)propan-2-yl)-1-(3′-fluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-1H-imidazol-4-yl)propan-2-ol.

In another embodiment, the present invention comprises a compound,isotope, or pharmaceutically acceptable salt thereof, which is2-(2-(2-(2,6-dichlorophenyl)propan-2-yl)-1-(3,3′-difluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-1H-imidazol-4-yl)propan-2-ol.

In another embodiment, the present invention comprises a compound,isotope, or pharmaceutically acceptable salt thereof, which is2-{2-[1-(2,6-dichlorophenyl)ethyl]-1-[3,3′-difluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl]-1H-imidazol-4-yl}propan-2-ol.

The various compounds described herein, or their pharmaceuticallyacceptable salts, may contain one or more asymmetric centers and maythus give rise to isomers, such as enantiomers, diastereomers, and otherstereoisomeric forms. Such forms may be defined, in terms of absolutestereochemistry, as (R)- or (S)-, or as (D)- or (L)- for amino acids.The present invention is meant to include all such possible individualstereoisomers and mixtures thereof, including their racemic andoptically pure enantiomeric or diastereomeric forms. The compounds arenormally prepared as racemates and can conveniently be used as such, oroptically active (+) and (−), (R)- and (S)-, or (D)- and (L)-isomers orcorresponding diastereomers may be prepared using chiral synthons orchiral reagents, or they may be resolved from racemic mixtures usingconventional techniques, such as chiral chromatography or reverse phaseHPLC. When the compounds described herein contain olefinic double bondsor other centers of geometric asymmetry, and unless specified otherwise,it is intended that the compounds include both E and Z geometricisomers.

The invention also includes isotopically-labeled compounds of theinvention, wherein one or more atoms is replaced by an atom having thesame atomic number, but an atomic mass or mass number different from theatomic mass or mass number usually found in nature. Examples of isotopessuitable for inclusion in the compounds of the invention includeisotopes of hydrogen, such as ²H or D and ³H or T, carbon such as ¹¹C,¹³C, and ¹⁴C, chlorine, such as ³⁶Cl, fluorine such as ¹⁸F, iodine, suchas ¹²³I and ¹²⁵I, nitrogen, such as ¹³N and ¹⁵N, oxygen, such as ¹⁵O,¹⁷O, and ¹⁸O, phosphorus, such as ³²P, and sulfur, such as ³⁵S. Certainisotopically-labeled compounds of the invention, for example, thoseincorporating a radioactive isotope, are useful in drug and/or substratetissue distribution studies. The radioactive isotopes tritium, ³H, andcarbon-14, ¹⁴C, are particularly useful for this purpose in view oftheir ease of incorporation and ready means of detection. Substitutionwith heavier isotopes such as deuterium, ²H or D, may afford certaintherapeutic advantages resulting from greater metabolic stability, forexample, increase in vivo half-life or reduced dosage requirements, andhence may be preferred in some circumstances. Substitution with positronemitting isotopes, such as ¹¹C, ¹⁸F, ¹⁵O, and ¹³N, can be useful inPositron Emission Topography (PET) studies for examining substratereceptor occupancy.

Isotopically-labeled compounds of the invention can generally beprepared by conventional techniques known to those skilled in the art orby processes analogous to those described herein, using an appropriateisotopically-labeled reagent in place of the non-labeled reagentotherwise employed.

DEFINITIONS

The following terms and expressions used herein have the indicatedmeanings.

“Nuclear receptor” refers to a receptor that activates or repressestranscription of one or more genes in the nucleus (but can also havesecond messenger signaling actions), typically in conjunction with othertranscription factors. The nuclear receptor is activated by the naturalcognate ligand for the receptor. Nuclear receptors are ordinarily foundin the cytoplasm or nucleus, rather than being membrane-bound. A nuclearreceptor is a member of a superfamily of regulatory proteins that arereceptors for various endogenous small molecules, e.g., steroids,retinoids, vitamin D and thyroid hormones. These proteins bind tocis-acting elements in the promoters of their target genes and modulategene expression in response to a ligand. Nuclear receptors may beclassified based on their DNA binding properties. For example, theglucocorticoid, estrogen, androgen, progestin and mineralocorticoidreceptors bind as homodimers to hormone response elements (HREs)organized as inverted repeats. Another example are receptors, includingthose activated by retinoic acid, thyroid hormone, vitamin D₃, fattyacids/peroxisome proliferators and ecdysone, that bind to HREs asheterodimers with a common partner, the retinoid X receptor (RXR). Amongthe latter receptors is LXR.

“Liver X receptor” or “LXR” refers to a nuclear receptor implicated incholesterol biosynthesis. As used herein, the term LXR refers to bothLXR_(α) and LXR_(β), two forms of the protein found in mammals. Liver Xreceptor-α or LXR_(α) refers to the receptor described in U.S. Pat. Nos.5,571,696, 5,696,233 and 5,710,004, and Willy et al. (1995) Gene Dev.9(9):1033-1045. Liver X receptor-β or LXR_(β) refers to the receptordescribed in Peet et al. (1998) Curr. Opin. Genet. Dev. 8(5):571-575;Song et al. (1995) Ann. N.Y. Acad. Sci. 761:38-49; Alberti et al. (2000)Gene 243(1-2):93-103; and references cited therein; and in U.S. Pat.Nos. 5,571,696, 5,696,233 and 5,710,004.

“Pharmaceutically acceptable” refers to those compounds, materials,compositions, and/or dosage forms which are, within the scope of soundmedical judgment, suitable for contact with the tissues of human beingsand animals without excessive toxicity, irritation, allergic response,or other problems or complications commensurate with a reasonablebenefit/risk ratio or which have otherwise been approved by the UnitedStates Food and Drug Administration as being acceptable for use inhumans or domestic animals.

“Pharmaceutically acceptable salt” refers to both acid and base additionsalts.

“Pharmaceutically acceptable acid addition salt” refers to those saltswhich retain the biological effectiveness and properties of the freebases, which are not biologically or otherwise undesirable, and whichare formed with inorganic acids such as hydrochloric acid, hydrobromicacid, sulfuric acid, nitric acid, phosphoric acid and the like, andorganic acids such as acetic acid, trifluoroacetic acid, propionic acid,glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid,succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid,cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid,p-toluenesulfonic acid, salicylic acid, and the like.

“Base addition salt” refers to those salts which retain the biologicaleffectiveness and properties of the free acids, which are notbiologically or otherwise undesirable. These salts are prepared fromaddition of an inorganic base or an organic base to the free acid. Saltsderived from inorganic bases include, but are not limited to, thesodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc,copper, manganese, aluminum salts and the like. Preferred inorganicsalts are the ammonium, sodium, potassium, calcium, and magnesium salts.Salts derived from organic bases include, but are not limited to, saltsof primary, secondary, and tertiary amines, substituted amines includingnaturally occurring substituted amines, cyclic amines and basic ionexchange resins, such as isopropylamine, trimethylamine, diethylamine,triethylamine, tripropylamine, ethanolamine, 2-dimethylaminoethanol,2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine,caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine,glucosamine, methylglucamine, theobromine, purines, piperazine,piperidine, N-ethylpiperidine, polyamine resins and the like.Particularly preferred organic bases are isopropylamine, diethylamine,ethanolamine, trimethylamine, di cyclohexylamine, choline and caffeine.

“Therapeutically effective amount” refers to that amount of a compoundwhich, when administered to a subject, is sufficient to effect treatmentfor a disease or disorder described herein. The amount of a compoundwhich constitutes a “therapeutically effective amount” will varydepending on the compound, the disorder and its severity, and the age ofthe subject to be treated, but can be determined routinely by one ofordinary skill in the art.

“Modulating” or “modulate” refers to the treating, prevention,suppression, enhancement or induction of a function, condition ordisorder. For example, it is believed that the compounds of the presentinvention can modulate atherosclerosis by stimulating the removal ofcholesterol from atherosclerotic lesions in a human.

“Treating” or “treatment” as used herein covers the treatment of adisease or disorder described herein, in a subject, preferably a human,and includes:

-   -   i. inhibiting a disease or disorder, i.e., arresting its        development; or    -   ii. relieving a disease or disorder, i.e., causing regression of        the disorder.

“Subject” refers to a warm blooded animal such as a mammal, preferably ahuman, or a human child, which is afflicted with, or has the potentialto be afflicted with one or more diseases and disorders describedherein.

“Atherosclerosis” refers to a process whereby atherosclerotic plaquesform within the inner lining of the artery wall leading toatherosclerotic cardiovascular diseases. Atherosclerotic cardiovasculardiseases can be recognized and understood by physicians practicing inthe relevant fields of medicine, and include without limitation,restenosis, coronary heart disease (also known as coronary arterydisease or ischemic heart disease), cerebrovascular disease includingischemic stroke, multi-infarct dementia, and peripheral vessel disease,including intermittent claudication, and erectile dysfunction.

“Dyslipidemia” refers to abnormal levels of lipoproteins in blood plasmaincluding both depressed and/or elevated levels of lipoproteins (e.g.,elevated levels of Low Density Lipoprotein, (LDL), Very Low DensityLipoprotein (VLDL) and depressed levels of High Density Lipoprotein(HDL).

“EC₅₀” refers to a dosage, concentration or amount of a particular testcompound that elicits a dose-dependent response at 50% of maximalexpression of a particular response that is induced, provoked orpotentiated by the particular test compound.

“Cholesterol” refers to a steroid alcohol that is an essential componentof cell membranes and myelin sheaths and, as used herein, incorporatesits common usage. Cholesterol also serves as a precursor for steroidhormones and bile acids.

“Triglyceride(s)” or “TGs” refers to three fatty acid moleculesesterified to a glycerol molecule and serve to store fatty acids whichare used by muscle cells for energy production or are taken up andstored in adipose tissue.

“IC₅₀” refers to an amount, concentration or dosage of a particular testcompound that achieves a 50% inhibition of a maximal response, such asmodulation of nuclear receptor, including the LXR_(α) or LXR_(β)activity, in an assay that measures such response.

“LXR” or “LXRs” refers to both LXR_(α) and LXR_(β).

“LXR_(α)” (LXR alpha) refers to all mammalian forms of such receptorincluding, for example, alternative splice isoforms and naturallyoccurring isoforms. Representative LXR_(α) species include, withoutlimitation the rat (Genbank Accession NM_031627), mouse (GenbankAccession BC012646), and human (GenBank Accession No. U22662) forms ofthe receptor.

“LXR_(β)” (LXR beta) refers to all mammalian forms of such receptorincluding, for example, alternative splice isoforms and naturallyoccurring isoforms. Representative LXR_(β) species include, withoutlimitation the rat (GenBank Accession NM_031626), mouse (GenbankAccession NM_009473), and human (GenBank Accession No. U07132) forms ofthe receptor.

“Obese” and “obesity” refer to a Body Mass Index (BMI) greater than 27.8kg/m² for men and 27.3 kg/m² for women (BMI equals weight(kg)/(height)²(m²).

Utility

The compounds of the invention exhibit valuable pharmacologicalproperties and are particularly useful as LXR agonists, antagonists,inverse agonists, partial agonists and antagonists, or are selective toLXR_(α) or to LXR_(β). The compounds of the invention are useful for thetreatment of diseases or disorders described herein, such as thoseassociated with, or having symptoms arising from the complications of,altered cholesterol transport, reverse cholesterol transport, fatty acidmetabolism, cholesterol absorption, cholesterol re-absorption,cholesterol secretion, cholesterol excretion, or cholesterol metabolism.

These diseases include, for example, atherosclerosis, atheroscleroticcardiovascular diseases, (see, e.g., International Patent ApplicationPublication Nos. WO 00/57915 and WO 00/37077), dyslipidemia,hyperglycemia, insulin resistance, diabetes, obesity, syndrome X (USPatent Application Publication No. 20030073614, International PatentApplication Publication No. WO 01/82917), excess lipid deposition inperipheral tissues such as skin (xanthomas) (see, e.g., U.S. Pat. Nos.6,184,215 and 6,187,814), stroke, peripheral occlusive disease, memoryloss (Brain Research (1997), Vol. 752, pp. 189-196), optic nerve andretinal pathologies (i.e., macular degeneration, retinitis pigmentosa),repair of traumatic damage to the central or peripheral nervous system(Trends in Neurosciences (1994), Vol. 17, pp. 525-530), prevention ofthe degenerative process due to aging (American Journal of Pathology(1997), Vol. 151, pp. 1371-1377), or Alzheimer's disease (see, e.g.,International Patent Application Publication No. WO 00/17334; Trends inNeurosciences (1994), Vol. 17, pp. 525-530), prevention of degenerativeneuropathies occurring in diseases such as diabetic neuropathies (see,e.g., International Patent Application Publication No. WO 01/82917),multiple sclerosis (Annals of Clinical Biochem. (1996), Vol. 33, No. 2,pp. 148-150), and autoimmune diseases (J. Lipid Res. (1998), Vol. 39,pp. 1740-1743).

Also provided, are methods of increasing the expression of ATP-BindingCassette (ABCA1), (see, e.g., International Patent ApplicationPublication No. WO 00/78972) thereby increasing reverse cholesteroltransport in mammalian cells using the claimed compounds andcompositions.

Accordingly in another aspect, the invention also includes methods toremove cholesterol from tissue deposits such as atherosclerotic plaquesor xanthomas in a subject with atherosclerosis or atheroscleroticcardiovascular disease manifest by clinical signs of such disease,wherein the methods comprise administering to the subject atherapeutically effective amount of a compound or composition of thepresent invention. Additionally, the instant invention also provides amethod for preventing or reducing the risk of a first or subsequentoccurrence of an atherosclerotic cardiovascular disease event includingischemic heart disease, ischemic stroke, multi-infarct dementia, andintermittent claudication comprising the administration of aprophylactically effective amount of a compound or composition of thepresent invention to a subject at risk for such an event.

The compounds of the present invention can also be used in methods fordecreasing hyperglycemia and insulin resistance, i.e., in methods fortreating diabetes (International Patent Application Publication No. WO01/82917), and in methods of treatment, prevention, or amelioration ofdisorders related to, or arising as complications of diabetes,hyperglycemia or insulin resistance including the cluster of diseasestates, conditions or disorders that make up “Syndrome X” (See US PatentApplication 20030073614) comprising the administration of atherapeutically effective amount of a compound or composition of thepresent invention to a subject in need of such treatment. Additionally,the instant invention also provides a method for preventing or reducingthe risk of developing hyperglycemia, insulin resistance, diabetes orsyndrome X in a subject, comprising the administration of aprophylactically effective amount of a compound or composition of thepresent invention to a subject at risk for such an event.

Diabetes mellitus, commonly called diabetes, refers to a disease processderived from multiple causative factors and characterized by elevatedlevels of plasma glucose, referred to as hyperglycemia. See, e.g.,LeRoith, D. et al., (eds.), DIABETES MELLITUS (Lippincott-RavenPublishers, Philadelphia, Pa. U.S.A. 1996). Uncontrolled hyperglycemiais associated with increased and premature mortality due to an increasedrisk for macrovascular diseases, including nephropathy, neuropathy,retinopathy, hypertension, cerebrovascular disease and coronary heartdisease. Therefore, control of glucose homeostasis is a criticallyimportant approach for the treatment of diabetes.

There are two major forms of diabetes: type 1 diabetes (formerlyreferred to as insulin-dependent diabetes or IDEM); and type 2 diabetes(formerly referred to as noninsulin dependent diabetes or NIDDM). Type 2diabetes is a disease characterized by insulin resistance accompanied byrelative, rather than absolute, insulin deficiency. Type 2 diabetes canrange from predominant insulin resistance with relative insulindeficiency to predominant insulin deficiency with some insulinresistance. Insulin resistance is the diminished ability of insulin toexert its biological action across a broad range of concentrations. Ininsulin resistant individuals, the body secretes abnormally high amountsof insulin to compensate for this defect. When inadequate amounts ofinsulin are present to compensate for insulin resistance and adequatecontrol of glucose, a state of impaired glucose tolerance develops. In asignificant number of individuals, insulin secretion declines furtherand the plasma glucose level rises, resulting in the clinical state ofdiabetes. Type 2 diabetes can be due to a profound resistance to insulinstimulating regulatory effects on glucose and lipid metabolism in themain insulin-sensitive tissues: muscle, liver and adipose tissue. Thisresistance to insulin responsiveness results in insufficient insulinactivation of glucose uptake, oxidation and storage in muscle andinadequate insulin repression of lipolysis in adipose tissue and ofglucose production and secretion in liver. In Type 2 diabetes, freefatty acid levels are often elevated in obese and some non-obesesubjects and lipid oxidation is increased.

Premature development of atherosclerosis and an increased rate ofcardiovascular and peripheral vascular diseases are characteristicfeatures of subjects with diabetes. Hyperlipidemia is an importantprecipitating factor for these diseases. Hyperlipidemia is a disordergenerally characterized by an abnormal increase in serum lipids, e.g.,cholesterol and triglyceride, in the bloodstream and is an importantrisk factor in developing atherosclerosis and heart disease. For areview of disorders of lipid metabolism, see, e.g., Wilson, J. et al.,(ed.), Disorders of Lipid Metabolism, Chapter 23, Textbook ofEndocrinology, 9th Edition, (W. B. Sanders Company, Philadelphia, Pa.U.S.A. 1998). Hyperlipidemia is usually classified as primary orsecondary hyperlipidemia. Primary hyperlipidemia is generally caused bygenetic defects, while secondary hyperlipidemia is generally caused byother factors, such as various disease states, drugs, and dietaryfactors. Alternatively, hyperlipidemia can result from both acombination of primary and secondary causes of hyperlipidemia. Elevatedcholesterol levels are associated with a number of disease states,including coronary artery disease, angina pectoris, carotid arterydisease, strokes, cerebral arteriosclerosis, and xanthoma.

Dyslipidemia, or abnormal levels of lipoproteins in blood plasma, is afrequent occurrence among diabetics, and has been shown to be one of themain contributors to the increased incidence of coronary events anddeaths among diabetic subjects (see, e.g., Joslin, E. Ann. Chim. Med.(1927), Vol. 5, pp. 1061-1079). Epidemiological studies since then haveconfirmed the association and have shown a several-fold increase incoronary deaths among diabetic subjects when compared with non-diabeticsubjects (see, e.g., Garcia, M. J. et al., Diabetes (1974), Vol. 23, pp.105-11 (1974); and Laakso, M. and Lehto, S., Diabetes Reviews (1997),Vol. 5, No. 4, pp. 294-315). Several lipoprotein abnormalities have beendescribed among diabetic subjects (Howard B., et al., Arteriosclerosis(1978), Vol. 30, pp. 153-162).

Further provided by this invention are methods of using the compounds ofthe invention to treat obesity, as well as the complications of obesity.Obesity is linked to a variety of medical disorders including diabetesand hyperlipidemia. Obesity is also a known risk factor for thedevelopment of type 2 diabetes (See, e.g., Barrett-Conner, E., Epidemol.Rev. (1989), Vol. 11, pp. 172-181; and Knowler, et al., Am. J Clin.Nutr. (1991), Vol. 53, pp. 1543-1551).

Administration and Formulation

A compound of the invention can be administered to subject in needthereof by any accepted route of administration. Acceptable routes ofadministration include, but are not limited to, buccal, cutaneous,endocervical, endosinusial, endotracheal, enteral, epidural,interstitial, intra-abdominal, intra-arterial, intrabronchial,intrabursal, intracerebral, intracisternal, intracoronary, intradermal,intraductal, intraduodenal, intradural, intraepidermal, intraesophageal,intragastric, intragingival, intraileal, intralymphatic, intramedullary,intrameningeal, intramuscular, intraovarian, intraperitoneal,intraprostatic, intrapulmonary, intrasinal, intraspinal, intrasynovial,intratesticular, intrathecal, intratubular, intratumor, intrauterine,intravascular, intravenous, nasal, nasogastric, oral, parenteral,percutaneous, peridural, rectal, respiratory (inhalation), subcutaneous,sublingual, submucosal, topical, transdermal, transmucosal,transtracheal, ureteral, urethral and vaginal.

A compound of the invention can be administered in any acceptable solid,semi-solid, liquid or gaseous dosage form. Acceptable dosage formsinclude, but are not limited to, aerosols, capsules, creams, emulsions,gases, gels, grains, liniments, lotions, ointments, pastes, powders,solutions, suspensions, syrups and tablets. Acceptable delivery systemsinclude, but are not limited to, biodegradable implants (e.g.,poly(DL-lactide), lactide/glycolide copolymers and lactide/caprolactonecopolymers), capsules, douches, enemas, inhalers, intrauterine devices,nebulizers, patches, pumps and suppositories.

A dosage form of the invention may be comprised solely of a compound ofthe invention or the compound of the invention may be formulated alongwith conventional excipients, pharmaceutical carriers, adjuvants, and/orother medicinal or pharmaceutical agents. Acceptable excipients include,but are not limited to, (a) antiadherents, such as croscarmellosesodium, crosprovidone, sodium starch glycolate, microcrystallinecellulose, starch and talc; (b) binders, such as cellulose, gelatin,hydroxypropyl cellulose, lactose, maltitol, polyethylene glycol,polyvinyl pyrrolidone, sorbitol, starch, sugar, sucrose and xylitol; (c)coatings, such as cellulose, shellac, zein and enteric agents; (d)disintegrants, such as cellulose, crosslinked polyvinyl pyrrolidone,sodium carboxymethyl cellulose, methylcellulose, microcrystallinecellulose, sodium starch glycolate and starch; (e) filling agents, suchas calcium carbonate, cellulose, dibasic calcium phosphate, glucose,lactose, mannitol, sorbitol and sucrose; (f) flavoring agents; (g)coloring agents; (h) glidants, such as calcium stearate, colloidalsilicon dioxide, glyceryl behenate, glyceryl monostearate, glycerylpalmitostearate, hydrogenated vegetable oil, magnesium stearate,magnesium trisilicate, mineral oil, polyethylene glycols, silicondioxide, starch, stearate, stearic acid, talc, sodium stearyl fumarate,sodium benzoate and zinc; (i) lubricants, such as calcium stearate,hydrogenated vegetable oils, magnesium stearate, mineral oil,polyethylene glycol, sodium stearyl fumarate, stearin, stearic acid andtalc; and (j) preservatives, such as chlorobutanol, citric acid,cysteine, methionine, methyl paraben, phenol, propyl paraben, retinylpalmitate, selenium, sodium citrate, sorbic acid, vitamin A, vitamin Cand vitamin E. Capsules may contain any of the afore listed excipients,and may additionally contain a semi-solid or liquid carrier, such as apolyethylene glycol or vegetable-based oils. Pharmaceutical carriersinclude soluble polymers, microparticles made of insoluble orbiodegradable natural and synthetic polymers, microcapsules ormicrospheres, lipoproteins, liposomes and micelles.

The pharmaceutical composition may be in the form of a liquid, e.g., anelixir, syrup, solution, emulsion, suspension, or other like forms ormay be presented as a dry product for reconstitution with water or othersuitable vehicle before use. Liquid preparations may containconventional additives such as (a) liquid diluents, such as water,saline, Ringer's solution, fixed oils such as synthetic mono ordiglycerides, or polyethylene glycols, glycerin, propylene glycol orother solvents; (b) surfactants, suspending agents, or emulsifyingagents, such as polyoxyethylene sorbitan fatty acid esters, saturatedpolyglycolized glycerides, monoglycerides, fatty acid esters, blockcopolymers of ethylene oxide and propylene oxide, polyoxyl stearates,ethoxylated castor oils, and ethoxylated hydroxystearic acids; (c)buffers, such as acetates, citrates or phosphates; (d) chelating agents,such as ethylenediaminetetraacetic acid; (e) antibacterial agents, suchas benzyl alcohol or methyl paraben; (f) antioxidants, such as ascorbicacid or sodium bisulfite; (g) isotonic agents, sodium chloride ordextrose; as well as sweetening and flavoring agents, dyes andpreservatives.

A pharmaceutical composition of the invention will contain atherapeutically effective amount of a compound of the invention, as anindividual stereoisomer or mixture of stereoisomers, or apharmaceutically acceptable salt thereof, with the remainder of thepharmaceutical composition comprised of one or more pharmaceuticallyacceptable excipients. Generally, for oral administration, a compound ofthe invention, as an individual stereoisomer or mixture ofstereoisomers, or a pharmaceutically acceptable salt thereof willcomprise from 1% to 99% by weight of a pharmaceutically acceptablecomposition, with the remainder of the composition comprised of one ormore pharmaceutically acceptable excipients. Typically, a compound ofthe invention, as an individual stereoisomer or mixture ofstereoisomers, or a pharmaceutically acceptable salt thereof willcomprise from 5% to 75% by weight of a pharmaceutically acceptablecomposition, with the remainder of the composition comprised of one ormore pharmaceutically acceptable excipients. For parenteraladministration, a compound of the invention, as an individualstereoisomer or mixture of stereoisomers, or a pharmaceuticallyacceptable salt thereof will comprise from 0.01% to 1% by weight of apharmaceutically acceptable composition. Methods for preparing thedosage forms of the invention are known, or will be apparent, to thoseskilled in this art; for example, see Remington's PharmaceuticalSciences, 18th Ed., (Mack Publishing Company, Easton, Pa., 1990).

A therapeutically effective amount of a compound of the invention willvary depending upon a sundry of factors including the activity,metabolic stability, rate of excretion and duration of action of thecompound, the age, weight, general health, sex, diet and species of thesubject, the mode and time of administration of the compound, thepresence of adjuvants or additional therapeutically active ingredientsin a composition, and the severity of the disease for which thetherapeutic effect is sought.

The compounds of the invention can be administered to human subjects atdosage levels in the range of about 0.1 to about 10,000 mg per day. Anormal human adult having a body weight of about 70 kilograms can beadministered a dosage in the range of from about 0.15 μg to about 150 mgper kilogram of body weight per day. Typically, a normal adult humanwill be administered from about 0.1 mg to about 25 mg, or 0.5 mg toabout 10 mg per kilogram of body weight per day. The compounds of theinvention may be administered in one or more unit dose forms. The unitdoses may be administered one to four times a day, or two times a day,or once a day. In an alternate method of describing an effective dose,an oral unit dose is one that is necessary to achieve a blood serumlevel of about 0.05 to 20 μg/ml or about 1 to 20 μg/ml in a subject. Theoptimum dose of a compound of the invention for a particular subject canbe determined by one of ordinary skill in the art.

Compounds of the invention, or an individual isomer or mixture ofisomers or a pharmaceutically acceptable salt thereof, may also beadministered simultaneously with, prior to, or after administration ofone or more of the therapeutic agents described below. Such combinationtherapy includes administration of a single pharmaceutical dosageformulation which contains a compound of the invention and one or moreadditional active agents, as well as administration of the compound ofthe invention and each active agent in its own separate pharmaceuticaldosage formulation. For example, a compound of the invention and anHMG-CoA reductase inhibitor can be administered to the subject togetherin a single oral dosage composition such as a tablet or capsule, or eachagent administered in separate oral dosage formulations. Where separatedosage formulations are used, the compounds of the invention and one ormore additional active agents can be administered at essentially thesame time, i.e., concurrently, or at separately staggered times, i.e.,sequentially; combination therapy is understood to include all theseregimens.

In one embodiment, the compounds of the invention are used incombination with one or more of the following therapeutic agents intreating atherosclerosis: antihyperlipidemic agents, plasma HDL-raisingagents, antihypercholesterolemic agents, cholesterol biosynthesisinhibitors (such as HMG CoA reductase inhibitors, such as lovastatin,simvastatin, pravastatin, fluvastatin, atorvastatin and rivastatin),acyl-coenzyme A:cholesterol acytransferase (ACAT) inhibitors, probucol,raloxifene, nicotinic acid, niacinamide, cholesterol absorptioninhibitors, bile acid sequestrants (such as anion exchange resins, orquaternary amines (e.g., cholestyramine or colestipol)), low densitylipoprotein receptor inducers, clofibrate, fenofibrate, benzofibrate,cipofibrate, gemfibrizol, vitamin B₆, vitamin B₁₂, anti-oxidantvitamins, β-blockers, anti-diabetes agents, angiotensin II antagonists,angiotensin converting enzyme inhibitors, platelet aggregationinhibitors, fibrinogen receptor antagonists, aspirin or fibric acidderivatives.

In another embodiment, the compounds of the invention are used incombination with one or more of the following therapeutic agents intreating cholesterol biosynthesis inhibitor, particularly an HMG-CoAreductase inhibitor. The term HMG-CoA reductase inhibitor is intended toinclude all pharmaceutically acceptable salt, ester, free acid andlactone forms of compounds which have HMG-CoA reductase inhibitoryactivity and, therefore, the use of such salts, esters, free acids andlactone forms is included within the scope of this invention. Compoundswhich have inhibitory activity for HMG-CoA reductase can be readilyidentified using assays well-known in the art. For instance, suitableassays are described or disclosed in U.S. Pat. No. 4,231,938 and WO84/02131. Examples of suitable HMG-CoA reductase inhibitors include, butare not limited to, lovastatin (MEVACOR®; see, U.S. Pat. No. 4,231,938);simvastatin (ZOCOR®; see, U.S. Pat. No. 4,444,784); pravastatin sodium(PRAVACHOL®; see, U.S. Pat. No. 4,346,227); fluvastatin sodium (LESCOL®;see, U.S. Pat. No. 5,354,772); atorvastatin calcium (LIPITOR®; see, U.S.Pat. No. 5,273,995) and rivastatin (also known as cerivastatin; see,U.S. Pat. No. 5,177,080). The structural formulae of these andadditional HMG-CoA reductase inhibitors that can be used in combinationwith the compounds of the invention are described at page 87 of M.Yalpani, “Cholesterol Lowering Drugs,” Chemistry & Industry, pp. 85-89(5 Feb. 1996). In presently preferred embodiments, the HMG-CoA reductaseinhibitor is selected from lovastatin and simvastatin.

In an additional embodiment, the compounds of the invention are used incombination with one or more of the following therapeutic agents intreating with one or more additional active diabetes agents depending onthe desired target therapy (see, e.g., Turner, N. et al., Prog. DrugRes. (1998), Vol. 51, pp. 33-94; Haffner, S., Diabetes Care (1998), Vol.21, pp. 160-178; and DeFronzo, R. et al. (eds.), Diabetes Reviews(1997), Vol. 5, No. 4). A number of studies have investigated thebenefits of combination therapies with oral agents (see, e.g., Mahler,R., J. Clin. Endocrinol. Metab. (1999), Vol. 84, pp. 1165-71; UnitedKingdom Prospective Diabetes Study Group: UKPDS 28, Diabetes Care(1998), Vol. 21, pp. 87-92; Bardin, C. W. (ed.), Current Therapy InEndocrinology And Metabolism, 6th Edition (Mosby—Year Book, Inc., St.Louis, Mo. 1997); Chiasson, J. et al., Ann. Intern. Med. (1994), Vol.121, pp. 928-935; Coniff, R. et al., Clin. Ther. (1997), Vol. 19, pp.16-26; Coniff, R. et al., Am. J. Med. (1995), Vol. 98, pp. 443-451;Iwamoto, Y. et al., Diabet. Med. (1996), Vol. 13, pp. 365-370;Kwiterovich, P., Am. J. Cardiol (1998), Vol. 82 (12A), pp. 3U-17U).These studies indicate that diabetes and hyperlipidemia modulation canbe further improved by the addition of a second agent to the therapeuticregimen.

In a further embodiment, the compounds of the invention are used incombination with one or more of the following therapeutic agents intreating diabetes: sulfonylureas (such as chlorpropamide, tolbutamide,acetohexamide, tolazamide, glyburide, gliclazide, glynase, glimepiride,and glipizide), biguanides (such as metformin), thiazolidinediones (suchas ciglitazone, pioglitazone, troglitazone, and rosiglitazone), andrelated insulin sensitizers, such as selective and non-selectiveactivators of PPARα, PPARβ and PPARγ; dehydroepiandrosterone (alsoreferred to as DHEA or its conjugated sulphate ester, DHEA-SO4);antiglucocorticoids; TNFα □inhibitors; α-glucosidase inhibitors (such asacarbose, miglitol, and voglibose), pramlintide (a synthetic analog ofthe human hormone amylin), other insulin secretogogues (such asrepaglinide, gliquidone, and nateglinide), insulin, as well as thetherapeutic agents discussed above for treating atherosclerosis.

In yet another embodiment, the compounds of the invention are used incombination with one or more of the following therapeutic agents intreating obesity or obesity-related disorders. Such agents, include, butare not limited to, phenylpropanolamine, phentermine, diethylpropion,mazindol, fenfluramine, dexfenfluramine, phentiramine, β₃ adrenoceptoragonist agents; sibutramine, gastrointestinal lipase inhibitors (such asorlistat), and leptins. Other agents used in treating obesity orobesity-related disorders include neuropeptide Y, enterostatin,cholecytokinin, bombesin, amylin, histamine H₃ receptors, dopamine D₂receptor modulators, melanocyte stimulating hormone, corticotrophinreleasing factor, galanin and gamma amino butyric acid (GABA).

Synthesis

The compounds of the present invention may be prepared in a number ofmethods well known to those skilled in the art, including, but notlimited to those described below, or through modifications of thesemethods by applying standard techniques known to those skilled in theart of organic synthesis. The compounds were named using ChemDraw Ultra9.0 or 10.0 (CambridgeSoft). The reagents and starting materials arecommercially available, or readily synthesized by well-known techniquesby one of ordinary skill in the arts. It is understood that in thefollowing description, combinations of substituents and/or variables ofthe depicted formulae are permissible only if such contributions resultin stable compounds. Unless otherwise indicated, all compoundsassociated with NMR and/or mass spectra data were prepared and the NMRand mass spectra measured.

In general, compounds of formula (0036) are prepared by first reactinganiline of formula (0032) with 2-(phenyl)-2-methylpropanenitrile (0031)in the presence of trimethylaluminum to give compounds of formula (0033)after standard isolation procedures (Scheme 1). In a subsequent step,exposure of amidine (0033) to a haloester, such as ethylα-bromopyruvate, under basic conditions at elevated temperature followedby dehydration conditions such as trifluoroacetic acid in ethanolprovides 1H-imidazole of formula (0034) after standard isolationprocedures. Compounds of formula (0034) are then subjected tofunctionality transformation, such as from ester to carbinol. In apalladium mediated coupling reaction, for example, Suzuki reactions,compounds of formula (0035) are then reacted with(2-fluoro-6-(sulfonyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)methanol(0017) (see Scheme 2 below) to afford compounds of formula (0036) afterstandard isolation procedures.

To a 500 mL round bottom flask attached with condenser was added4-bromo-2,6-difluorobenzoic acid (16.0 g, 67.5 mmol) and anhydrous THF(110 mL). The reaction flask was cooled in an ice bath prior to dropwiseaddition of 1.0 M lithium bis-(trimethylsilyl)amide (74 mL, 1.1 equiv).The reaction suspension was stirred at room temperature for 20 min priorto addition of sodium thiomethoxide (5.21 g, 74.2 mmol). The reactionsolution was allowed to stir at reflux for 3 hr. The reaction wasdetermined to be complete after quenching a reaction aliquot in diluteaq. HCl and running GCMS: found m/z=265, 267 parent ions. The cooledreaction mixture was quenched with H₂O and diluted with EtOAc (200 mL).The reaction mixture was transferred to a separatory funnel, and 1.0 Naq. HCl was added to give a pH=2-3 solution. The ethyl acetate layer wasseparated, washed with brine, dried over Na₂SO₄, and concentrated invacuo to afford 14.6 g (81% yield) of the intermediate6-fluoro-4-bromo-2-methylsulfanyl-benzoic acid as a waxy white solid. ¹HNMR (400 MHz, CDCl₃) δ 7.18 (s, 1H), 7.12 (dd, J=8 Hz, 1H), 2.49 (s,3H); GCMS m/z=265, 267 [M]⁺.

Alternatively, the intermediate 6-fluoro-4-bromo-2-methylsulfanyl-benzoic acid was prepared as follows:

To a 20 L flask was charged dimethyl formamide (14.5 L, 10.0 vol),followed by sodium hydroxide (293.7 g, 1.2 eq) and the reaction masscooled to −15 to −10° C. 4-bromo-2,6-difluorobenzoic acid (1450 g, 1.0equiv) was added over a period of 10-15 min at −15 to −10° C. andstirred for an additional 10-15 min. Sodium thiomethoxide (514.6 g, 1.2equiv) was added over a period of 5-10 min at −10 to −5° C. Oncompletion of the addition the temperature of the reaction was raised to25-28° C. over a period of 45 to 60 min and maintained at thattemperature 1.5-2 h. The temperature of the reaction was then raised to60-65° C. over a period of 30-60 min and maintained at 60-65° C. for 5 huntil the reaction was deemed complete. The reaction mixture was thencooled to 20-25° C. and quenched with a cooled (5-10° C.) solution of 2NHCl (5.045 L of 12N HCl in 30.3 L water). Following the quench, ethylacetate (14.5 L, 10 vol) was added and the mixture stirred for 10-15min. The phases were separated and the aqueous layer was extracted withethyl acetate (7.25 L, 5 vol). The two phases were separated and thecombined organic layer was washed with a brine solution (725 g of NaClin 3.625 L of water). The phases were separated and the organic layerwas washed with water (5.0 vol, 7.25 L). The phases were separated andthe organic layer was dried over sodium sulfate (1450 g). The organiclayer was filtered to remove the sodium sulfate, which was then washedwith ethyl acetate (2.9 L, 2 vol). The organic layer was concentratedunder reduced pressure at 45-50° C./30-40 mm Hg to ˜1 to 1.2 volumes andpetroleum ether (7.25 L, 5 vol) was added at 40-45° C. over a period of15-20 min. The solution was cooled to 20-25° C. over a period of 20-25min. The solid was filtered and washed with petroleum ether (2.9 L, 2.0vol) and the product dried under vacuum at 25-28° C., 0.4 to 0.7 mbar toafford 1410 g (87%, 99.40 Area %) of the intermediate6-fluoro-4-bromo-2-methylsulfanyl-benzoic acid.

Step 2b Preparation of (4-bromo-2-fluoro-6-(methylthio)phenyl)methanol

Into a N₂ purged 500 mL round bottom flask attached with condenser wasadded 6-fluoro-4-bromo-2-methylsulfanyl-benzoic acid (14.6 g, 55.0 mmol)and anhydrous THF (70 mL). The reaction solution was allowed to cool to0° C. prior to dropwise addition of a 1.0 M BH₃-THF (83 mL, 1.5 equiv)solution in THF. The reaction solution was stirred at room temperaturethen at reflux for an additional 2 hr. The reaction solution was cooledprior to quenching with a 1:1 H₂O/THF solution. The reaction solutionwas transferred to a separatory funnel with EtOAc (100 mL) and anaqueous solution of K₂CO₃ was added. The ethyl acetate phase wasseparated, washed with brine, dried over Na₂SO₄, and concentrated invacuo. The crude product was chromatographed through a 110 g SiO₂ columnusing a solvent gradient of 100% Hx to 55% EtOAc. The purified titleproduct was obtained as a solid white wax (13.7 g, 99% yield). ¹H NMR(400 MHz, CDCl₃) δ 7.13 (s, 1H), 7.06 (dd, J₁=8 Hz, J₂=2 Hz, 1H), 4.77(s, 2H), 2.51 (s, 3H), 2.20-2.05 (br s, 1H); GCMS m/z=251, 253 [M]⁺.

Alternatively, the intermediate(4-bromo-2-fluoro-6-(methylthio)phenyl)methanol was prepared as follows:

To a 20 L flask was charged 4-bromo-2-fluoro-6-(methylthio)benzoic acid(1400 g, 1.0 eq) followed by THF (14 L, 10 vol) under nitrogen. To thissolution was added borane-dimethyl sulfide complex (802.41 g, 1000 mL)at 25-28° C. over a period of 30-45 min. The reaction temperature wasraised to 60-65° C. over a period of 30-45 min and the temperaturemaintained until HPLC showed <1% of4-bromo-2-fluoro-6-(methylthio)benzoic acid (˜3-4 h). On completion ofthe reaction the mixture was cooled to 10-15° C. over a period of 30-40min. The reaction was then quenched with methanol (2.1 L, 1.5 vol) overa period of 1 to 1½ h at 10-15° C. The reaction mass was thenconcentrated under vacuum at 40-50° C./0.4 to 0.7 mbar to 1 to 1.5volumes. The resultant mixture was dissolved in DCM (8.4 L, 6 vol). Theorganic layer was washed with an ammonium chloride solution (560 g NH₄Clin 2.8 L water, 2 vol). The phases were separated and the organic layerwas washed with 10% NaHCO₃ solution (2.8 L, 2 vol), saturated brinesolution (2.1 L, 1.5 vol) and water (4.2 L, 3 vol). The organic layerwas separated and dried over sodium sulfate (700 g). The sodium sulfatewas removed by filtration and washed with DCM (2.8 L, 2 vol). Theorganic layer was concentrated under vacuum at 40-45° C./0.4 to 0.7 mbarto 1 to 1.2 vol to afford the product which was dried under vacuum at45-50° C./0.4 to 0.7 mbar. The title product was obtained in 90% yield(1200 g) with 90.07 Area %.

Step 2c Preparation of(4-bromo-2-fluoro-6-(methanesulfonyl)phenyl)-methanol

To a 500 mL flask was added(4-bromo-2-fluoro-6-(methylthio)phenyl)methanol (13.7 g, 54.6 mmol) andanhydrous dichloromethane (125 mL). The solution was cooled to 0-3° C.in an ice bath prior to portionwise addition of 3-chloroperbenzoic acid(77% max., Aldrich) (18.8 g, 2 equiv). The reaction solution was thenallowed to warm to room temperature where it remained for 18 h. Thereaction was then concentrated in vacuo to remove dichloromethane andthe residue was washed into a separatory funnel with ethyl acetate and 1M aq. NaOH. The ethyl acetate layer was separated, washed with 1 M aq.NaOH, dried over Na₂SO₄, and concentrated in vacuo. The residue waspurified by flash chromatography (Biotage, 65×200 mm SiO₂ column,gradient elution from 100% hexanes to 90% ethyl acetate). Appropriatefractions were combined and concentrated in vacuo to afford the titlecompound as a colorless, semi-crystalline solid, yield: 8.1 g (52%). ¹HNMR (400 MHz, DMSO-d₆) δ 7.98 (dd, J=8 Hz, 1H), 7.91 (s, 1H), 5.45 (t,J=8 Hz, 1H), 4.88 (dd, J, =8 Hz, =2 Hz, 2H), 3.42 (s, 3H); ¹⁹F NMR (400MHz, DMSO-d₆) δ −111.8 ppm; GCMS m/z=283, 285 [M]⁺.

Step 2d Preparation of(2-fluoro-6-(methylsulfonyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)methanol

To a 100 mL round bottom flask, purged with dry N₂, was weighed(4-bromo-2-fluoro-6-(methanesulfonyl)phenyl)-methanol (1.98 g, 6.99mmol), bis(pinacolato)diboron (2.13 g 1.2 equiv),dichloro[1,1′-bis(diphenylphosphino) ferrocene]palladium (II)dichloromethane adduct (560 mg, 10 mol %), potassium carbonate (2.06 g,3 equiv), and DMSO (25 mL). The resulting suspension was allowed to stirat 90° C. for 3 h. An aliquot of reaction solution was found to containno more starting bromide as determined by LCMS analysis. The cooledreaction suspension was diluted with ethyl acetate (50 mL) and water (50mL) and filtered through a Celite padded Buchner Funnel. The resultingfiltrate was transferred to a separatory funnel, and the organic phasewas separated. The aqueous phase was extracted with ethyl acetate, andthe combined ethyl acetate phases were washed with brine, dried overNa₂SO₄, and concentrated in vacuo. The residue was purified by silicagel flash chromatography (Biotage SP-1, 40 g SiO₂ column, gradientelution from 100% hexanes to 60% ethyl acetate) to afford a clearviscous oil. The product was isolated as an amorphous white powder bydissolving in dichloromethane and reprecipitation resulted upon additionof hexanes. The title compound was isolated as a solid white powder,yield: 1.9 g (82% yield). ¹H NMR (400 MHz, CDCl₃) δ 8.28 (s, 1H), 7.79(d, J=8 Hz, 1H), 5.03 (d, J=8 Hz, 2H), 3.23 (s, 3H) 3.05 (t, J=8 Hz,1H), 1.35 (s, 6H); ¹⁹F NMR (400 MHz, CDCl₃) δ −116.3 ppm.

Alternatively, the intermediate(2-fluoro-6-(methylsulfonyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)methanolwas prepared as follows:

To a 500 mL jacketed reactor equipped with a stir bar, temperatureprobe, reflux condenser and a nitrogen inlet was charged methyltetrahydroran (MeTHF) (75 mL, 5 volumes) followed by potassium acetate(5.2 g, 52.98 mmoles, 1 equiv.) and(oxydi-2,1-phenylene)bis(diphenylphosphine) (322 mg; 597.3 μmoles,0.01125 equiv.) and bis(pinacolato)diboron (17.51 g, 68.95 mmoles, 1.3equiv.). The reaction flask was evacuated to less than 150 Torr, andthen back filled with nitrogen. This degassing procedure was repeated 3times. Pd(OAc)₂ (94.2 mg; 419.6 μmoles, 0.0075 equiv.) was charged tothe reactor and the reaction flask was evacuated to less than 150 Torr,and then back filled with nitrogen and the sequence repeated 3 times.The resulting slurry was allowed to age at 20-25° C. for 15 min. Uponcompletion of the 15 min age, the slurry was heated to an internaltemperature of 80° C. As the mixture in the reactor was heating totemperature, in a separate flask was charged(4-bromo-2-fluoro-6-(methanesulfonyl)phenyl)-methanol (15.03 g, 53.09mmoles, 1 equiv.) followed by MeTHF (75 ml, 5 volumes). The resultingsolution was degassed by bubbling nitrogen subsurface for at least 15min. prior to use. Once the catalyst mixture had reached reflux, thedegassed solution of(4-bromo-2-fluoro-6-(methanesulfonyl)phenyl)-methanol in MeTHF was addedto the reaction in a single portion and allowed to react. The reactiontypically takes ˜20 hours to complete after the addition of substrate.Upon completion (typically <0.75 RAP of starting material the reactionwas cooled to 20-25° C. Once at RT the reaction was diluted with MeTHF(75 ml, 5 volumes) and washed with a 5 wt % NaCl solution (7.5 volumes,110 ml) for at least 15 min. The phases were separated and the upperproduct rich MeTHF stream was filtered through Celite to removeinsoluble palladium residues. The Celite cake was washed with MeTHF (75ml, 5 volumes). The reaction was treated with functionalized silica (30equiv) to remove palladium and color. The slurry was agitated for atleast 60 min and then filtered to remove the silica. The used silica waswashed with MeTHF (5 volumes, 75 ml). The combined organic phase waswashed with water (5 volumes, 75 ml). The organic was distilled to 5volumes (75 ml) under vacuum (60-70 Torr, bath temp of 30° C.). When the75 ml landmark was reached the distillation was stopped and heptane (75ml, 5 volumes) was added drop wise to the reaction solution. After ˜35ml of heptanes had been added the product began to crystallize from thesolution. On completion of the addition the product was isolated byfiltration and the wet cake washed with MeTHF-heptanes (1:9) solution(2×75 ml) and dried at 50° C. The title product was obtained a whitesolid, 13.64 g, (78% yield) with 99.58 Area %.

Example 12-(1-(3-chloro-3′-fluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-2-(2-(2-fluorophenyl)propan-2-yl)-1H-imidazol-4-yl)propan-2-ol

Example 1a Preparation of 2-(2-fluorophenyl)-2-methylpropanenitrile

To a 500 mL 3-neck round bottom flask with an attached addition funnelthat has been purged with dry N₂, was added 2-fluorophenylacetonitrile(11.0 g, 81.4 mmol) and anhydrous THF (70 mL). The reaction solution wascooled to −10° C. prior to dropwise addition of a 1.0 M potassiumtert-butoxide solution (195 mL, 2.4 molar equiv) in THF. The reactionsolution was stirred at −10° C. for 20 min prior to addition ofiodomethane (15.2 mL, 244 mmol). The reaction solution was allowed tostir warming to room temperature for 4 hr. The reaction solution wasquenched by addition of aq NH₄Cl and diluted with EtOAc (200 mL). Theorganic phase was partitioned, washed with aq NH₄Cl, dried over Na₂SO₄,filtered, concentrated in vacuo and chromatographed through a 240 g SiO₂column on the Biotage SP-1 using a solvent gradient of 100% Hx to 50%EtOAc to afford 10.1 g (76% yield) of title product. GCMS m/z=163 [M]⁺.

Example 1b Preparation ofN-(4-bromophenyl)-2-(2-fluorophenyl)-2-methylpropanimidamide

To an oven dried, N₂ purged 250 mL round bottom flask attached withaddition funnel was added 4-bromoaniline (7.31 g, 42.5 mmol) andanhydrous toluene (40 mL). To the reaction solution at 0° C. was added a2.0 M Me₃Al (32 mL, 1.5 molar equiv) solution. The reaction solution wasstirred at 0° C. for 30 min, then a solution of2-(2-fluorophenyl)-2-methylpropanenitrile (7.62 g, 46.7 mmol) in toluene(25 mL) was added to the reaction flask. The reaction solution wasallowed to stir at 90° C. for 5 hr. The cooled reaction solution wasquenched with an aq sodium potassium tartrate solution. After standing20 min, the organic phase was partitioned and washed with sodiumpotassium tartrate solution. The organic solution was extracted with 1Naq HCl (100 mL×3). The combined aq HCl solution was neutralized byaddition of 1N aq NaOH and extracted with dichloromethane (200 mL×2).The dichloromethane product solution was dried over Na₂SO₄, filtered,and concentrated in vacuo to afford the title compound (5.5 g, 39%yield). GCMS m/z=334, 336 [M]⁺.

Example 1c Preparation of ethyl1-(4-bromophenyl)-2-(2-(2-fluorophenyl)propan-2-yl)-1H-imidazole-4-carboxylate

To a 250 mL round bottom flask attached with condenser was addedN-(4-bromophenyl)-2-(2-fluorophenyl)-2-methylpropanimidamide (5.0 g, 15mmol), anhydrous THF (80 mL), NaHCO₃ (2.52 g, 30 mmol), and 90% ethylbromopyruvate (1.90 mL, 15.1 mmol). The reaction mixture was stirred at70° C. for 2 hr prior to analysis by LCMS. The cooled reaction mixturewas decanted and concentrated in vacuo. The residue was taken intotoluene (65 mL) and acetic acid (1.8 mL). The solution was stirred atreflux for 1 hr. The cooled solution was washed with H₂O (150 mL×3),dried over Na₂SO₄, filtered, concentrated in vacuo, and chromatographedthrough a SiO₂ column using a 100% Hx to 70% EtOAc gradient to affordpurified title compound (4.3 g, 67% yield). LCMS (ES): m/z=431.3, 433.3[M+H]⁺.

Example 1d Preparation of2-(1-(4-bromophenyl)-2-(2-(2-fluorophenyl)propan-2-yl)-1H-imidazol-4-yl)propan-2-ol

To a 250 mL round bottom flask, purged with dry N₂ and attached withaddition funnel, was added a 3.0M MeMgBr (12 mL, 3.7 equiv) solution inEt₂O. The flask was cooled to 0° C. prior to dropwise addition of ethyl1-(4-bromophenyl)-2-(2-(2-fluorophenyl)propan-2-yl)-1H-imidazole-4-carboxylate(4.22 g, 9.78 mmol) in a solution of anhydrous dichloromethane (80 mL).The reaction solution was allowed to stir, warming to room temperatureover 1 hr. The reaction solution was quenched by addition of aq NH₄Cl.The mixture was poured to a separatory funnel and the dichloromethanelayer was partitioned, dried over Na₂SO₄, filtered, concentrated andchromatographed through a 40 g SiO₂ column using a gradient of 100% Hxto 70% EtOAc to yield the title compound (3.19 g, 78% yield). LCMS (ES):m/z=417.3, 419.3 [M+H]⁺.

Example 1 Preparation of2-(1-(3′-fluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-2-(2-(2-fluorophenyl)propan-2-yl)-1H-imidazol-4-yl)propan-2-ol

To a 50 mL round bottom flask was added2-(1-(4-bromophenyl)-2-(2-(2-fluorophenyl)propan-2-yl)-1H-imidazol-4-yl)propan-2-ol(380 mg, 911 μmol), DME (25 mL) and H₂O (6 mL). The solution was spargedwith N₂ for 10 min prior to addition of(2-fluoro-6-(methylsulfonyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)methanol(360 mg, 1.09 mmol), potassium carbonate (380 mg, 2.73 mmol), anddichloro[1,1′-bis (diphenylphosphino)ferrocene]palladium (II)dichloromethane adduct (74 mg, 91 μmol). The reaction mixture wasallowed to stir at 80° C. for 2 h. The cooled reaction solution wasdiluted with EtOAc (30 mL) and filtered through a Celite padded Buchnerfunnel. The filtrate was washed with aq NH₄Cl (150 mL×2). The organicphase was dried over Na₂SO₄, filtered, and concentrated in vacuo. Theresidue was purified by silica gel flash chromatography (Biotage SP-1,25 g SiO₂ column, gradient elution from 5% EtOAc to 100% EtOAc) toafford the title compound (100 mg, 20% yield). ¹H-NMR (400 MHz, CDCl₃) δ8.02 (d, J=2 Hz, 1H), 7.51 (dd, J₁₌₂ Hz, J₂=10 Hz, 1H), 7.29 (d, J=92H), 7.08-7.16 (mult, 1H), 6.85-6.92 (mult, 3H), 6.77-6.84 (mult, 2H),6.65 (s, 1H), 5.09 (d, J=6 Hz, 2H), 3.35 (s, 1H), 3.30 (2, 3H), 3.02 (t,J=6 Hz, 1H), 1.72 (s, 6H), 1.62 (s, 6H); ¹⁹F NMR (400 MHz, CDCl₃) δ−112.1, −113.5 ppm; LCMS (ES) m/z=541.3 [M+H]⁺, 563.2 [M+Na]⁺.

Examples 2-8

All of the following compounds were made in a similar manner to thatdescribed in Example 1 using appropriate anilines and2-(phenyl)-2-methylpropanenitriles. If not commercially available,nitriles were made using standard techniques that are readily apparentto one skilled in the art.

No. Name Structure Data 2 2-(2-(2-(2-chloro-6-fluorophenyl)propan-2-yl)-1-(3′- fluoro-4′-(hydroxymethyl)-3- methyl-5′-(methylsulfonyl)biphenyl-4-yl)- 1H-imidazol-4-yl)propan-2-ol

MS (ES): 589.3 [M + H]⁺ 3 2-(1-(3-chloro-3′-fluoro-4′-(hydroxymethyl)-5′- (methylsulfonyl)biphenyl-4-yl)-2-(2-(2,6-dichlorophenyl)propan-2- yl)-1H-imidazol-4-yl)propan-2-ol

MS (ES): 627.2 [M + H]⁺ 4 2-(2-(2-(2-chloro-3-fluorophenyl)propan-2-yl)-1-(3′- fluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)- 1H-imidazol-4-yl)propan-2-ol

MS (ES): 575.3 [M + H]⁺ 5 2-(2-(2-(2,6-dichlorophenyl)propan-2-yl)-1-(3′- fluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)- 1H-imidazol-4-yl)propan-2-ol

MS (ES): 591.5 [M + H]⁺ 6 2-(2-(2-(2-Chloro-phenyl)propan-2-yl)-1-(3,3′-difluoro-4′- hydroxymethyl-5′-(methylsulfonyl)biphenyl-4-yl)- 1H-imidazol-4-yl)-propan-2-ol

MS (ES): 575.3 [M + H]⁺ 7 2-(2-(2-(2-chloro-6-fluorophenyl)propan-2-yl)-1-(3,3′- difluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)- 1H-imidazol-4-yl)propan-2-ol

MS (ES) 593.3, 595.3 [M + H]⁺ 8 2-{1-(3,3′-Difluoro-4′-hydroxymethyl-5′- methanesulfonyl-biphenyl-4-yl)-2-[2-(2-fluorophenyl)propan-2-yl]- 1H-imidazol-4-yl}-propan-2-ol

MS (ES): 559.2 [M + H]⁺

Compound 2 has the following NMR characteristics: 1H NMR (400 MHz,CDCl₃) δ 8.02 (s, 1H), 7.56-7.49 (m, 1H), 7.35 (d, J=2.0, 1H), 7.12-6.96(m, 3H), 6.68 (d, J=8.2, 1H), 6.66-6.60 (m, 1H), 6.56 (s, 1H), 5.08 (d,J=5.4, 2H), 3.36 (s, 1H), 3.29 (s, 3H), 2.92 (t, J=7.0, 1H), 2.07 (s,3H), 1.97 (d, J=2.4, 3H), 1.72 (d, J=7.4, 3H), 1.59 (s, 6H).

Compound 3 has the following NMR characteristics: 1H NMR (400 MHz,CDCl₃) δ 8.00 (m, 1H), 7.57 (d, J=2.1, 1H), 7.55-7.49 (m, 1H), 7.13 (s,1H), 7.11 (s, 1H), 7.07 (dd, J=8.3, 2.1, 1H), 7.01-6.95 (m, 1H), 6.81(d, J=8.3, 1H), 6.59 (s, 1H), 5.09 (d, J=5.4, 2H), 3.30 (s, 3H), 3.26(m, 1H), 2.89 (t, J=7.0, 1H), 2.06 (s, 3H), 1.92 (s, 3H), 1.61 (s, 3H),1.59 (s, 3H).

Compound 4 has the following NMR characteristics: 1H NMR (400 MHz,CDCl₃) δ 7.97 (s, 1H), 7.47 (dd, J=10.0, 1.8, 1H), 7.31-7.20 (m, 2H),7.00 (d, J=8.3, 2H), 6.92-6.71 (m, 3H), 6.65 (s, 1H), 5.08 (dd, J=7.0,1.6, 2H), 3.29 (s, 3H), 3.27 (s, 1H), 2.90 (t, J=7.0, 1H), 1.82 (s, 6H),1.60 (s, 6H).

Compound 5 has the following NMR characteristics: 1H-NMR (400 MHz,DMSO-d6) δ 7.89-7.90 (mult, 1H), 7.82-7.85 (mult, 1H), 7.52 (d, J=8.6Hz, 2H), 7.16 (d, J=8.6 Hz 2H), 7.07-7.09 (mult, 2H), 6.94-6.98 (mult,1H), 6.80 (s, 1H), 5.55 (t, J=5.2 Hz, 1H), 4.93-4.95 (mult, 2H), 4.65(s, 1H), 3.45 (s, 3H), 1.96 (s, 6H), 1.45 (s, 6H).

Compound 6 has the following NMR characteristics: 1H NMR (400 MHz,CDCl₃) δ 7.97 (s, 1H), 7.46 (dd, J=9.9, 1.8, 1H), 7.23-7.18 (m, 1H),7.12 (dd, J=10.3, 1.9, 1H), 6.97 (ddd, J=23.4, 9.0, 4.0, 2H), 6.88-6.79(m, 2H), 6.61 (s, 1H), 5.08 (d, J=5.4, 2H), 3.30 (s, 3H), 3.27-3.23 (m,1H), 2.92 (t, J=6.9, 1H), 1.61 (s, 12H).

Compound 7 has the following NMR characteristics: 1H-NMR (DMSO-d6, 400MHz) δ 7.95-7.90 (m, 2H), 7.62 (dd, 1H, J=11, 1.5 Hz), 7.33 (dd, 1H,J=9.5, 1.5 Hz), 7.13-7.08 (m, 3H), 6.85 (s, 1H), 6.80-6.70 (m, 1H), 5.57(t, 1H, J=5.3 Hz), 4.95 (d, 2H, J=4.3 Hz), 4.71 (s, 1H), 3.47 (s, 3H),1.85 (s, 6H), 1.46 (s, 6H).

Compound 8 has the following NMR characteristics: 1H-NMR (DMSO-d6, 400MHz) δ 7.96-7.90 (m, 2H), 7.49 (dd, 1H, J=11, 1.5 Hz), 7.34 (dd, 1H,J=9.5, 1.5 Hz), 7.20-7.10 (m, 1H), 7.05-6.94 (m, 2H), 6.90-6.75 (m, 3H),5.57 (t, 1H, J=5.3 Hz), 4.94 (d, 2H, J=4.3 Hz), 4.70 (s, 1H), 3.47 (s,3H), 1.68 (s, 6H), 1.47 (s, 6H).

Example 92-(2-(2-(2,6-dichlorophenyl)propan-2-yl)-1-(3,3′-difluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-1H-imidazol-4-yl)propan-2-ol

Example 9a Preparation of 2-(2,6-dichlorophenyl)-2-methylpropanenitrile

To a 1 M solution of potassium tert-butoxide (403 mL, 403 mmol) at −66°C. (acetone/dry ice) was slowly added 2-(2,6-dichlorophenyl)acetonitrile(25.0 g, 134 mmol) in anhydrous THF (150 mL). The mixture was stirred at−66° C. for 20 minutes. Then, iodomethane (33.6 mL, 538 mmol) was addeddrop-wise over 25 minutes at −66° C. At this stage, it was exothermicand a large amount of light yellow precipitate was observed. Thesuspension was stirred at −60° C. for 30 minutes. The reaction mixturewas quenched with 200 mL ice water, and extracted with ether (3×150 mL).The organics were combined, washed with 150 mL brine, dried over Na₂SO₄,and concentrated on a rotary evaporator. The crude product (30 g, yellowoil) was purified by column chromatography (ISCO, 330 g silica, 20%EtOAc in hexanes) to afford2-(2,6-dichlorophenyl)-2-methylpropanenitrile (28.2 g, 132 mmol, 98%yield) as a light yellowish oil. ¹H-NMR (CDCl₃, 400 MHz) δ 7.35 (d, 2H,J=8.03 Hz), 7.16 (t, 1H, J=8.0 Hz), 2.09 (s, 6H); ¹³C-NMR (CDCl₃, 126MHz) δ 134.6, 133.8, 131.4, 129.0, 124.1, 38.6, 29.2; MS m/e 214.10(M+H⁺); HPLC (XBridge 5μ C18 4.6×50 mm, 4 mL/min, Solvent A: 10%MeOH/water with 0.2% H₃PO₄, Solvent B: 90% MeOH/water with 0.2% H₃PO₄,gradient with 0-100% B over 4 minutes): 3.16 minutes.

Example 9b Preparation ofN-(4-bromo-2-fluorophenyl)-2-(2,6-dichlorophenyl)-2-methylpropanimidamide

2-(2,6-Dichlorophenyl)-2-methylpropanenitrile (20 g, 93 mmol) and4-bromo-2-fluoroaniline (28.4 g, 149 mmol) were dissolved in anhydrouso-xylene (200 mL) and heated to 100° C. under N₂. Trimethylaluminum (2M) in toluene (140 mL, 280 mmol) was added drop-wise (˜0.9 mL perminute) over 2.5 hours while the reaction mixture was stirred at 100° C.After addition, the reaction mixture was stirred at 100° C. for 30minutes, and then cooled to −5° C. The reaction mixture was verycarefully quenched with potassium sodium tartrate (20 g in 100 mL water)(Caution: gas and heat formation). The reaction mixture was filteredthrough Celite 545. The filtrate was washed with 1N HCl (4×70 mL). Theaqueous was neutralized with 2N NaOH and extracted with EtOAc (4×100mL). The organics were combined, washed with brine, dried with Na₂SO₄,and concentrated on a rotary evaporator to afford 24 g of crude product.The crude product was recrystallized with 72 mL of MTBE and 240 mL ofhexane to giveN-(4-bromo-2-fluorophenyl)-2-(2,6-dichlorophenyl)-2-methylpropanimidamide(17.5 g, 43.3 mmol, 46.4% yield) as a white solid (purity: 99%). ¹H-NMR(MeOD, 400 MHz) δ 7.42 (d, 2H, J=8.0 Hz), 7.30 (m, 2H), 7.16 (t, 1H,J=8.0 Hz), 6.93 (t, 1H, J=8.0 Hz), 2.11 (s, 6H); ¹³C-NMR (DMSO-d₆, 100MHz) δ 166.5, 156.1, 153.7, 140.6, 138.5, 135.9, 131.4, 128.6, 128.0,125.7, 119.5, 112.9, 50.0, 29.2; MS m/e 403.09 (M+H⁺); HPLC (XBridge 5μC18 4.6×50 mm, 4 mL/min, Solvent A: 10% MeOH/water with 0.2% H₃PO₄,Solvent B: 90% MeOH/water with 0.2% H₃PO₄, gradient with 0-100% B over 4minutes): 2.32 minutes.

Example 9c Preparation of ethyl1-(4-bromo-2-fluorophenyl)-2-(2-(2,6-dichlorophenyl)propan-2-yl)-4-hydroxy-4,5-dihydro-1H-imidazole-4-carboxylate

To a mixture ofN-(4-bromo-2-fluorophenyl)-2-(2,6-dichlorophenyl)-2-methylpropanimidamide(48.0 g, 119 mmol), K₂CO₃ (41.0 g, 297 mmol) in toluene (180 mL) and THF(180 mL) at 55° C. was added slowly a solution of ethyl3-bromo-2-oxopropanoate (23.3 mL, 166 mmol) in 24 mL of THF over 50minutes. The reaction mixture was kept at 55° C. for 1.5 hours. A whiteslurry was observed. The reaction mixture was cooled to 5° C. HCl (0.5N,450 mL) was added drop-wise (end point pH=9˜10). After addition, thesuspension was cooled to 0° C. The solid was collected by filtration,washed with water (2×50 mL), and then dried in a vacuum oven at 60° C.overnight. Ethyl1-(4-bromo-2-fluorophenyl)-2-(2-(2,6-dichlorophenyl)propan-2-yl)-4-hydroxy-4,5-dihydro-1H-imidazole-4-carboxylate(59 g, 114 mmol, 96% yield) was obtained as a white solid. ¹H-NMR(CDCl₃, 400 MHz) δ 7.11 (m, 3H), 6.96 (m, 2H), 6.72 (t, 1H, J=8.28 Hz),4.35 (m, 2H), 4.25 (d, 1H, J=10.5 Hz), 3.80 (d, 1H, J=10.8 Hz), 1.98 (s,3H), 1.93 (s, 3H), 1.38 (t, 3H, J=7.03 Hz); ¹³C-NMR (CDCl₃, 126 MHz) δ173.0, 171.5, 159.8, 157.8, 137.3, 135.7, 132.1, 131.1, 128.1, 127.4,125.6, 122.2, 120.1, 93.5, 62.5, 45.5, 30.2, 14.0; MS m/e 517.05 (M+H⁺);HPLC (XBridge 5μ C18 4.6×50 mm, 4 mL/min, Solvent A: 10% MeOH/water with0.2% H₃PO₄, Solvent B: 90% MeOH/water with 0.2% H₃PO₄, gradient with0-100% B over 4 minutes): 2.74 minutes.

Example 9d Preparation of ethyl1-(4-bromo-2-fluorophenyl)-2-(2-(2,6-dichlorophenyl)propan-2-yl)-1H-imidazole-4-carboxylate

To a mixture of ethyl1-(4-bromo-2-fluorophenyl)-2-(2-(2,6-dichlorophenyl)propan-2-yl)-4-hydroxy-4,5-dihydro-1H-imidazole-4-carboxylate(38 g, 73 mmol) in EtOH (200 mL) was added TFA (25.0 g, 220 mmol). Themixture was subsequently heated to 95° C. HPLC analysis after 2.5 hoursshowed <1% of alcohol intermediate remaining. The mixture was dilutedwith 300 mL of CH₂Cl₂ and cooled to approximately 5° C. with an icebath. The mixture was neutralized with 1N NaOH (120 mL) and the organiclayer was separated. The aqueous layer was extracted with CH₂Cl₂ (2×100mL). The combined organic layers were concentrated on a rotaryevaporator to give crude material. Recrystallization in EtOH (5 mL/1 g)provided 32 g of ethyl1-(4-bromo-2-fluorophenyl)-2-(2-(2,6-dichlorophenyl)propan-2-yl)-1H-imidazole-4-carboxylateas an off-white solid (86% yield). ¹H-NMR (DMSO-d₆, 400 MHz) δ 7.92 (s,1H), 7.16 (d, 1H, J=8.0 Hz), 7.22 (m, 3H), 7.11 (m, 1H), 7.04 (t, 1H,J=12.0 Hz), 4.25 (q, 2H, J=8.0 Hz), 1.94 (s, 6H), 1.27 (t, 3H, J=8.0Hz); MS m/e 502.68 (M+H⁺); HPLC (XBridge C18 4.6×50 mm, 4 mL/min,Solvent A: 10% MeOH/water with 0.2% H₃PO₄, Solvent B: 90% MeOH/waterwith 0.2% H₃PO₄, gradient with 0-100% B over 4 minutes): 3.87 minutes.

Example 9e Preparation of2-(1-(4-bromo-2-fluorophenyl)-2-(2-(2,6-dichlorophenyl)propan-2-yl)-1H-imidazol-4-yl)propan-2-ol

To a mixture of methylmagnesium bromide (60.0 mL, 180 mmol, 3M in ether)in 120 mL of THF cooled with an ice/salt bath (˜15 to −17° C.) was addedslowly a solution of ethyl 1-(4-bromo-2-fluorophenyl)-2-(242,6-dichlorophenyl)propan-2-yl)-1H-imidazole-4-carboxylate (30 g, 60 mmol)in 65 mL of CH₂Cl₂ and 87 mL of THF over 45 minutes. The internaltemperature was carefully kept below 0° C. A further 2×20 mL of CH₂Cl₂was used to wash forward the residual material. The reaction mixturetemperature was maintained below 0° C. for 1 hour with stirring. Thenthe reaction mixture was diluted with 100 mL of CH₂Cl₂, and saturatedNH₄Cl was added slowly. The resulting mixture was extracted with CH₂Cl₂(2×80 mL). Organics were combined, washed with brine, dried with Na₂SO₄,and concentrated on a rotary evaporator to afford2-(1-(4-bromo-2-fluorophenyl)-2-(2-(2,6-dichlorophenyl)propan-2-yl)-1H-imidazol-4-yl)propan-2-ol(28.5 g, 58.6 mmol, 98% yield) as a white solid. ¹H-NMR (CDCl₃, 400 MHz)δ 7.13 (dd, 1H, J=9.03, 2.01 Hz), 7.09 (s, 1H), 7.07 (s, 1H), 6.93 (m,2H), 6.75 (t, 1H, J=8.16 Hz), 6.55 (s, 1H), 3.18 (s, 1H), 2.00 (s, 6H),1.58 (s, 6H); ¹³C-NMR (CDCl₃, 126 MHz) δ 158.1, 156.1, 154.5, 147.8,139.3, 135.7, 131.3, 130.3, 127.8, 126.9, 122.7, 119.8, 115.1, 68.7,44.8, 31.1, 29.9; MS m/e 485.05 (M+H⁺); HPLC (XBridge 5μ C18 4.6×50 mm,4 mL/min, Solvent A: 10% MeOH/water with 0.2% H₃PO₄, Solvent B: 90%MeOH/water with 0.2% H₃PO₄, gradient with 0-100% B over 4 minutes): 2.78minutes.

Example 9 Preparation of2-(2-(2-(2,6-dichlorophenyl)propan-2-yl)-1-(3,3′-difluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-1H-imidazol-4-yl)propan-2-ol

To a 1 L 3-necked round bottom flask under nitrogen was added2-(1-(4-bromo-2-fluorophenyl)-2-(2-(2,6-dichlorophenyl)propan-2-yl)-1H-imidazol-4-yl)propan-2-ol(12.0 g, 24.7 mmol),[2-fluoro-6-methanesulfonyl-4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-methanol(9.78 g, 29.6 mmol), K₂CO₃ (10.2 g, 74 mmol), DME (120 mL) and water (12mL). The mixture was heated to 60° C., and then1,1′-bis(diphenylphosphino)ferrocene palladium (II) chloride complex(4.06 g, 4.94 mmol) was added under nitrogen. The reaction mixture washeated to 80° C. for 30 minutes. The resulting darkly colored mixturewas cooled with an ice bath, and partitioned in 200 mL of CH₂Cl₂ and 200mL of water. The organic layers were combined and dried with Na₂SO₄.After concentration, the crude product was purified by flashchromatography (ISCO, 330 g silica, 0% to 100% EtOAc in hexanes) toafford 12.79 g of crude product (85% yield) as a light yellow solid.

Recrystallization was carried out by dissolving 9.5 g of crude productin acetone (80 mL) at 65° C. The resulting solution was cooled slowly to25° C. over 5 hours, and then cooled to 0° C. for an additional 30minutes. Crystals began to form at 45° C. The solid was collected byfiltration and rinsed with cold acetone. After drying in an oven at 45°C. under vacuum for 14 hours, 4.9 g of pure product was obtained. Torecover additional crystalline product, the mother liquid wasconcentrated to approximately 10 mL and passed through a silica pad.EtOAc (100 mL) was used to elute the compound. The filtrate wasconcentrated under vacuum to give a crude solid. The crude solid wasrecrystallized in acetone following the procedure above to afford anadditional 2.5 g of product. The combined recovery for the two cropsafter recrystallization was a 78% yield. ¹H-NMR (DMSO-d₆, 400 MHz) δ7.94 (m, 2H), 7.63 (dd, 1H, J=11.29, 1.51 Hz), 7.34 (d, 1H, J=9.54 Hz),7.14 (m, 3H), 7.05 (m, 1H), 6.83 (s, 1H), 5.58 (t, 2H, J=5.27 Hz), 4.96(d, 2H, J=4.27 Hz), 4.70 (s, 1H), 3.46 (s, 3H), 1.96 (s, 6H), 1.45 (s,6H); MS m/e 609.16 (M+H⁺); HPLC (XBridge 5μ, C18 4.6×50 mm, 4 mL/min,Solvent A: 10% MeOH/water with 0.2% H₃PO₄, Solvent B: 90% MeOH/waterwith 0.2% H₃PO₄, gradient with 0-100% B over 4 minutes): 2.56 minutes.

Alternatively, Example 9 was prepared as follows:

To a 1 L 3-necked round bottom flask under nitrogen was addedmethyltetrahydrofuran (“MeTHF”, 6.9 kg),2-(1-(4-bromo-2-fluorophenyl)-2-(2-(2,6-dichlorophenyl)propan-2-yl)-1H-imidazol-4-yl)propan-2-ol(1.994 kg, 4.1 moles) and(2-fluoro-6-(methylsulfonyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)methanol(1.38 kg, 4.19 moles). The mixture was agitated at 23° C. for 15 minuntil all the solids dissolved. At the conclusion of this period,(oxydi-2,1-phenylene)bis(diphenylphosphine) (0.022 kg, 0.041 moles) andPd(OAc)₂ (0.01 kg, 0.045 moles) were added as a slurry via a subsurfaceline. Upon completion of addition, the mixture was rinsed withadditional MeTHF (1.65 kg). The resulting mixture was evacuated to lessthan 80 Torr and backfilled with nitrogen. This process was repeated twomore times. After completion of the degassing sequence, the reactionmixture was agitated for at least 15 min and a clear, golden color wasobserved. In a separate reaction vessel, a solution of potassiumhydroxide (0.352 kg) in water (10.00 kg) was prepared and degassed bysparging the solution with nitrogen gas for at least 15 min prior touse. The KOH solution (10.35 kg) was transferred into the reactor byvacuum. The reaction temperature exhibited a known exotherm from 20° C.to 29° C. Upon completion of addition, the resulting biphasic mixturewas degassed by a series of pressure swings. The mixture was warmed tobetween 45-50° C. where it was stirred for at least 2 h. After thistime, the reaction mixture was analyzed by HPLC, which indicated thereaction was complete. The reaction mixture was cooled to 23° C. and thestirring was stopped. The mixture was allowed to separate for 30 min andthe lower spent KOH stream was removed. The product rich organic waspassed through a column of thiourea functionalized silica gel (0.782 kg)(Silicycle) at ˜0.1 kg per min to remove the palladium. The product richorganic phase was washed with a 5% NaHCO₃ solution (5 vol) and thephases separated. The organic phase was washed with water (5 vol) andthe organic and aqueous phases separated.

The product rich organic phase was polish filtered into a clean reactionvessel and then concentrated to ˜8 volumes (˜16 L) under vacuum (80Torr, Tjacket=60° C.). Once at the prescribed volume, the reactionmixture was allowed to cool to 25° C. Once at the prescribed temperaturethe reaction mixture was seeded with2-(2-(2-(2,6-dichlorophenyl)propan-2-yl)-1-(3,3′-difluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-1H-imidazol-4-yl)propan-2-ol(0.5%, 0.008 kg). The resulting slurry was stirred at 25° C. for about18 h. At the conclusion of this period, the reaction mixture wasconcentrated to ˜8 L under vacuum (150 Torr, Tjacket=60° C.). Once atthe prescribed volume, the reaction mixture was heated to 50° C. andisopropyl acetate (IPAc, 13.90 kg) was added to the reactor during a 90min period. Upon completion of addition, the reaction mixture was cooledto 25° C. during a 3 h period. Once at the prescribed temperature thereaction mixture was stirred at room temperature for about 16 h. At theconclusion of this period, the reaction mixture was filtered,deliquored, and washed with additional IPAc (10.4 kg). The filter cakewas dried via suction on the filter under a stream of dry nitrogen toyield a white solid. The white solid was transferred to a dryer anddried at 50° C. under full vacuum to afford 2.03 kg of product (81%yield, 99.40 AP, 98 wt %).

Example 102-(2-(2-(2,6-dichlorophenyl)propan-2-yl)-1-(3,3′-difluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-1H-imidazol-4-yl)[(¹³CD₃)₂]propan-2-ol

Preparation of2-(1-(4-bromo-2-fluorophenyl)-2-(2-(2,6-dichlorophenyl)propan-2-yl)-1H-imidazol-4-yl[(¹³CD₃)₂]propan-2-ol

Oven-dried magnesium (86 mg, 3.52 mmol) and anhydrous diethyl ether(3.20 mL) were transferred to an oven-dried 25 mL 14/20 round bottomflask under argon. [¹³CD₃]-Iodomethane (467 mg, 3.20 mmol) was added atroom temperature and stirred at 33° C. for 1 hour. Visual signsindicated that the Grignard reagent formed (clear suspension changed tocloudy mixture with frothing and exotherm). The solution was cooled toroom temperature and transferred via cannula to a chilled solution(ice/water bath) of ethyl1-(4-bromo-2-fluorophenyl)-2-(2-(2,6-dichlorophenyl)propan-2-yl)-1H-imidazole-4-carboxylate(400 mg, 0.800 mmol) in anhydrous dichloromethane (2.60 mL) under argon.The flask in which the Grignard reagent was prepared was rinsed withanhydrous ether (2×400 μL) and transferred via cannula to the ethylester-containing flask. The reaction mixture was allowed to slowly warmto room temperature and stirred for 1 hour. HPLC analysis indicated<0.3% of starting material was present. The reaction was cooled to 0°C., diluted with anhydrous dichloromethane (800 μL) and quenched by theslow, careful addition of sat. aqueous ammonium chloride (8 mL). The twolayers were separated, and the aqueous layer was extracted withdichloromethane (3×4 mL). The combined organic extracts wereconcentrated in vacuo to obtain 443.5 mg of crude product as a whitesolid. The crude product described above was combined with 244.3 mg(white solid) obtained from a similar reaction. The combined product waspurified by silica gel flash chromatography (Isco RediSep cartridge, 12g) and eluted with 10 to 20% EtOAc in hexane. 30 mL fractions werecollected. The fractions were checked by TLC (Silica, 50% EtOAc, 50%hexane, R_(f)=0.41) and HPLC. The pure product-containing fractions werecombined and concentrated in vacuo to yield 531.2 mg of product as awhite solid (90% yield): ¹H-NMR (400 MHz, CD₃OD) δ ppm: 6.47-7.58 (m,7H), 2.01 (br s, 6H). HPLC: (YMC ODS-AQ, 3 μm, 150×4.6 mm, Mobile PhaseA=0.05% TFA in H₂O, Mobile Phase B=0.05% TFA in ACN, 0 min 50% B, 9 min95% B, 15 min 95% B, 15.5 min 50% B. Flow rate=1 ml/min) T_(r)=9.23 min(at 220 nm, Chemical purity=98.8%), LCMS (+ion) m/z=487 (0%), 493 (59%),495 (100%), 497 (47%), 498 (8%).

Preparation of2-(2-(2-(2,6-dichlorophenyl)propan-2-yl)-1-(3,3′-difluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-1H-imidazol-4-yl)[(¹³CD₃)₂]propan-2-ol

To a 25 mL 14/20 round bottom flask under argon was added2-(1-(4-bromo-2-fluorophenyl)-2-(2-(2,6-dichlorophenyl)propan-2-yl)-1H-imidazol-4-yl)[(¹³CD₃)₂]propan-2-ol(0.283 g, 0.572 mmol),(2-fluoro-6-(methylsulfonyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)methanol(0.227 g, 0.686 mmol),1,1′-Bis(diphenylphosphino)-ferrocene-palladium(II)dichloridedichloromethane complex (0.094 g, 0.114 mmol), potassium carbonate(0.237 g, 1.716 mmol), DME (4.30 ml) and water (0.215 ml) that had beenpreviously sparged with argon. The mixture was heated to 80° C. for 1hour. HPLC and LCMS analysis indicated starting material had beenconsumed. The reaction mixture was cooled with an ice-water bath,partitioned between dichloromethane (10 mL) and water (10 mL). Theaqueous layer was extracted with dichloromethane (3×10 mL). The combinedorganic extracts were concentrated in vacuo to give 643.6 mg as a darksemi-solid.

Previously, a similar reaction for the preparation of2-(2-(2-(2,6-dichlorophenyl)propan-2-yl)-1-(3,3′-difluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-1H-imidazol-4-yl)[(¹³CD₃)₂]propan-2-ol had been completed that afforded 462.3 mg of adark solid. The crude products from both experiments were combined andpurified by silica gel flash chromatography (Isco RediSep cartridge, 80g) after dissolving in dichloromethane and pre-absorbing onto silicagel. The flash column was eluted with 25-50% EtOAc in hexane. 30 mLfractions were collected. Fractions were checked by TLC (Silica, 50%EtOAc, 50% hexane, R_(f)=0.09) and HPLC before combining the pureproduct-containing fractions and concentrating in vacuo to give 468.3 mgof2-(2-(2-(2,6-dichlorophenyl)propan-2-yl)-1-(3,3′-difluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-1H-imidazol-4-yl)[(¹³CD₃)₂]propan-2-olas a yellow solid.

This material was further purified by recrystallization in acetone (4mL) at 65° C., cooled slowly to 25° C. over 5 hours (crystals started toform at 40° C.) then cooled to 0° C. for an additional 30 minutes. Thesolid was collected by filtration, rinsed with cold acetone and dried invacuo to give 66.2 mg of the title compound as an off-white solid.

The mother liquor from this first recrystallization was concentrated andthe residue was recrystallized from a minimal amount of acetone (1 mL)using the same procedure as outlined above to obtain a second crop of276.4 mg of crystalline product as an off-white solid.

The mother liquor from the second recrystallization was concentrated andpurified by preparative HPLC, T_(r)=15.0 min (Prep HPLC conditions:Synergi Hydro-RP column, 4μ, 80 Å, 21.2×250 mm, Mobile Phase A=H₂O,Mobile Phase B=ACN, 0 min 30% B, 25 min 100% B. Flow rate=16.0 ml/min.UV at 220 nm.)

The 3 purified isolates were combined to give 401.1 mg of2-(2-(2-(2,6-dichlorophenyl)propan-2-yl)-1-(3,3′-difluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-1H-imidazol-4-yl)[(¹³CD₃)₂]propan-2-ol as a pale yellow solid (64% yield). ¹H-NMR (400MHz, DMSO-d₆)) δ ppm: 7.86-7.96 (m, 2H), 7.62 (dd, J=11.33 Hz, 2.01 Hz,1H), 7.33 (dd, J=8.31 Hz, 1.76 Hz, 1H), 7.09-7.19 (m, 3H), 6.99-7.09 (m,1H), 6.81 (s, 1H), 5.53-5.62 (m, 1H), 4.89-4.99 (m, 2H), 4.67 (t, J=3.15Hz, 1H), 3.45 (s, 3H), 2.08 (residual acetone, 8 mol %), 1.95 (s, 6H).¹³C-NMR (400 MHz, D6-DMSO) 29.61 (t, J=109.88 Hz)

HPLC: (YMC ODS-AQ, 3 pin, 150×4.6 mm, Mobile Phase A=0.05% TFA in H₂O,Mobile Phase B=0.05% TFA in ACN, 0 min 50% B, 9 min 95% B, 15 min 95% B,15.5 min 50% B. Flow rate=1 ml/min) T_(r)=9.66 min (at 220 nm, Chemicalpurity=99.8%). LCMS (+ion) m/z=609 (0%), 617 (100%), 618 (31%), 619(74%), 620 (22%), 621 (16%), 622 (4.3%), 623 (1.3%).

Examples 11-20

The following compounds were made in a similar manner to that describedin Example 1, by substituting various phenyl acetonitrile reagents inplace of 2-(phenyl)-2-methylpropanenitriles as the starting material:

No. Name Structure Data 11 2-(2-(2,4-dichlorobenzyl)-1-(3,3′-difluoro-4′- (hydroxymethyl)-5′- (methylsulfonyl)biphenyl-4-yl)-1H-imidazol-4-yl)propan- 2-ol

MS (ES): 581.3, 583.3 [M + H]⁺ 12 2-(1-(3,3′-difluoro-4′-(hydroxymethyl)-5′- (methylsulfonyl)biphenyl-4- yl)-2-(2-(trifluoromethyl)benzyl)-1H- imidazol-4-yl)propan-2-ol

MS (ES): 581.3 [M + H]⁺ 13 2-(1-(3-chloro-3′-fluoro-4′-(hydroxymethyl)-5′- (methylsulfonyl)biphenyl-4- yl)-2-(2-chloro-4-fluorobenzyl)-1H-imidazol-4- yl)propan-2-ol

MS (ES): 581.3, 583.3[M + H]⁺ 14 2-(2-(2-chloro-4-fluorobenzyl)-1-(3,3′-difluoro- 4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4- yl)-1H-imidazol-4-yl)propan- 2-ol

MS (ES): 565.3 [M + H]⁺ 15 2-(2-(2,4-dichlorobenzyl)-1-(3′-fluoro-4′-(hydroxymethyl)- 5′-(methylsulfonyl)biphenyl-4-yl)-1H-imidazol-4-yl)propan- 2-ol

MS (ES): 563.2, 565.2 [M + H]⁺ 16 2-(1-(3,3′-difluoro-4′-(hydroxymethyl)-5′- (methylsulfonyl)biphenyl-4-yl)-2-(2-fluorobenzyl)-1H- imidazol-4-yl)propan-2-ol

MS (ES): 531.2 [M + H]⁺ 17 2-(1-(3′-fluoro-4′- (hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4- yl)-2-(2-methylbenzyl)-1H-imidazol-4-yl)propan-2-ol

MS (ES): 509.5 [M + H]⁺; 531.2 [M + Na]⁺ 18 2-(2-(2,6-dichlorobenzyl)-1-(3′-fluoro-4′-(hydroxymethyl)- 5′-(methylsulfonyl)biphenyl-4-yl)-1H-imidazol-4-yl)propan- 2-ol

MS (ES): 586.5 [M + H]⁺ 19 2-[2-(2-Chloro-5-fluoro-benzyl)-1-(3′-fluoro-4′- hydroxymethyl-5′- methanesulfonyl-biphenyl-4-yl)-1H-imidazol-4-yl]-propan- 2-ol

MS (ES): 547.3 [M + H]⁺ 20 2-[2-(2-Chloro-benzyl)-1-(3,3′-difluoro-4′-hydroxymethyl-5′- methanesulfonyl-biphenyl-4-yl)-1H-imidazol-4-yl]-propan- 2-ol

MS (ES): 547.3 [M + H]⁺

Example 21 Example for Compound 21

Into a 1 L flask was weighed 25.0 g (134 mmol) of2,6-dichlorophenylacetonitrile and 250 mL of anhydrous THF. Theresulting solution was cooled to −70° C. and 134 mL of 1.0 M potassiumtert-butoxide (1.0 M) in THF was added followed by 8.4 mL of iodomethane(1.0 eq). The reaction was stirred at −70° C. for 1 h then was allowedto warm to room temperature over 3 h. The reaction was concentrated invacuo to remove THF then was washed into a separatory funnel with ethylacetate and 1 M HCl. The ethyl acetate was separated, washed withbisulfite, brine, was dried (Na₂SO₄), and concentrated in vacuo. Theresidue was purified by silica gel flash chromatography (Biotage, 300 gSiO₂, gradient elution from 100% hexanes to 10% ethyl acetate over 1 h).Appropriate fractions were combined and concentrated in vacuo to affordthe desired product as a colorless oil, ˜99% pure by GC, yield: 12.2 g(45%). ¹H NMR (CDCl₃, 400 MHz) δ 7.36 (d, J=8 Hz, 2H), 7.22 (t, J=8 Hz,1H), 4.84 (q, J=7 Hz, 1H), 1.07 (d, J=7 Hz, 3H).

Compound 21 was made in a similar matter to that described in Example 1using appropriate aniline and 2-(phenyl)propanenitrile as startingmaterial.

No. Name Structure Data 21 2-{2-[1-(2,6-dichlorophenyl)ethyl]-1-[3,3′-difluoro-4′-(hydroxymethyl)- 5′-(methylsulfonyl)biphenyl-4-yl]-1H-imidazol-4-yl}propan-2-ol

Compound 11 has the following NMR characteristics: 1H NMR (400 MHz,CDCl₃) δ 8.10 (d, J=1.2, 1H), 7.59 (dd, J=9.9, 1.8, 1H), 7.47-7.37 (m,2H), 7.24 (dd, J=5.3, 3.2, 2H), 7.13 (dd, J=8.3, 2.1, 1H), 7.05 (d,J=8.4, 1H), 6.89 (s, 1H), 5.10 (d, J=4.4, 2H), 4.09 (s, 2H), 3.30 (s,3H), 3.25 (d, J=17.4, 1H), 2.86 (s, 1H), 1.63 (s, 6H).

Compound 12 has the following NMR characteristics: 1H NMR (400 MHz,DMSO) δ 8.06 (d, J=8.7, 2H), 7.99-7.86 (m, 1H), 7.70 (d, J=8.3, 1H),7.58 (m, 3H), 7.40 (t, J=7.6, 1H), 7.21 (d, J=7.7, 1H), 7.13 (s, 1H),5.57 (t, J=5.3, 1H), 4.95 (d, J=4.7, 2H), 4.81 (s, 1H), 4.13 (s, 2H),3.45 (s, 3H), 1.46 (s, 6H).

Compound 13 has the following NMR characteristics: 1H NMR (400 MHz,CDCl₃) δ 8.10 (s, 1H), 7.71 (d, J=2.0, 1H), 7.60 (dd, J=9.9, 1.8, 1H),7.49 (dd, J=8.2, 2.1, 1H), 7.22 (d, J=8.2, 1H), 7.12 (dd, J=8.6, 6.1,1H), 6.96 (dd, J=8.5, 2.6, 1H), 6.89-6.78 (m, 2H), 5.10 (s, 2H), 4.05(s, 2H), 3.31 (s, 3H), 3.00 (s, 1H), 2.77 (s, 1H), 1.63 (2, J=5.5, 6H).

Compound 14 has the following NMR characteristics: 1H NMR (400 MHz,DMSO) δ 7.85 (t, J=4.5, 2H), 7.70 (dd, J=11.4, 1.8, 1H), 7.50 (dd,J=8.3, 1.7, 1H), 7.35 (t, J=8.2, 1H), 7.09 (dd, J=8.8, 2.6, 1H),7.03-6.80 (m, 3H), 5.35 (t, J=5.3, 1H), 4.73 (d, J=4.1, 2H), 4.56 (s,1H), 3.80 (s, 2H), 3.38 (t, J=6.4, 1H), 3.23 (s, 3H), 1.21 (s, 6H).

Compound 15 has the following NMR characteristics: 1H NMR (400 MHz,CDCl₃) δ 8.11 (d, J=1.2, 1H), 7.72-7.53 (m, 3H), 7.28 (dd, J=18.0, 4.9,5H), 7.15 (dd, J=8.3, 2.1, 1H), 7.05 (d, J=8.4, 1H), 6.96 (s, 1H), 5.09(s, 2H), 4.12 (s, 2H), 3.30 (s, 3H), 3.28 (s, 1H), 2.93 (s, 1H), 1.64(s, 6H).

Compound 16 has the following NMR characteristics: 1H NMR (400 MHz,CDCl₃) δ 8.09 (s, 1H), 7.60 (dd, J=9.9, 1.8, 1H), 7.39 (m, 2H), 7.23 (t,J=8, 1H), 7.18-7.06 (m, 2H), 7.00 (m, 1H), 6.89-6.81 (m, 2H), 5.10 (dd,J=7.0, 1.6, 2H), 4.06 (s, 2H), 3.30 (s, 3H), 2.92 (t, J=7.0, 1H), 2.70(s, 1H), 1.64 (s, 6H).

Compound 17 has the following NMR characteristics: 1H NMR (400 MHz,CDCl₃) δ 8.10 (s, 1H), 7.64-7.55 (m, 3H), 7.25 (m, 2H), 7.16-7.00 (m,3H), 6.92 (s, 1H), 6.81 (d, J=7.5, 1H), 5.08 (dd, J=7.1, 1.7, 2H), 4.02(s, 2H), 3.28 (s, 3H), 2.90 (t, J=7.1, 1H), 2.76 (s, 1H), 2.16 (d,J=8.6, 3H), 1.64 (s, 6H).

Compound 18 has the following NMR characteristics: 1H NMR (400 MHz,CDCl₃) δ 8.12 (s, 1H), 7.70-7.59 (m, 3H), 7.45 (d, J=8.4, 2H), 7.24 (d,J=8.0, 2H), 7.16-7.07 (m, 1H), 6.85 (s, 1H), 5.10 (d, J=5.6, 2H), 4.31(s, 2H), 3.30 (s, 3H), 3.07 (s, 1H), 2.94 (t, J=7.0, 1H), 1.55 (s, 6H).

Compound 19 has the following NMR characteristics: 1H NMR (400 MHz,CDCl₃) δ 8.11 (d, J=1.1, 1H), 7.62 (ddd, J=12.0, 8.4, 1.8, 3H),7.28-7.26 (m, 3H), 6.97 (s, 1H), 6.93-6.76 (m, 2H), 5.09 (s, 2H), 4.14(s, 2H), 3.29 (br s, 4H), 2.91 (s, 1H), 1.65 (s, 6H).

Compound 20 has the following NMR characteristics: 1H NMR (400 MHz,CDCl₃) δ 8.08 (d, J=1.1, 1H), 7.57 (dd, J=9.9, 1.8, 1H), 7.38 (ddd,J=10.3, 9.2, 2.0, 2H), 7.23-7.17 (m, 2H), 7.18-7.01 (m, 3H), 6.89 (d,J=0.7, 1H), 5.09 (s, 2H), 4.14 (s, 2H), 3.37 (s, 1H), 3.30 (s, 3H), 2.92(s, 1H), 1.64 (s, 6H).

Compound 21 has the following NMR characteristics: 1H NMR (400 MHz,CDCl₃) δ 7.97 (s, 1H), 7.47 (dd, J=9.8, 1.6, 1H), 7.18 (d, J=10.6, 1H),7.12-7.04 (m, 2H), 6.96 (d, J=7.9, 2H), 6.90-6.81 (m, 1H), 6.79 (s, 1H),5.13 (s, 2H), 4.94 (q, J=7.0, 1H), 3.82 (s, 1H), 3.39 (d, J=24.5, 1H),3.36 (s, 3H), 1.81 (d, J=7.1, 3H), 1.63 (s, 6H).

Standard physiological, pharmacological and biochemical procedures areavailable for testing the compounds to identify those that possessbiological activities that modulate the activity of the LXRs (LXR_(α)and LXR_(β)). Such assays include, for example, biochemical assays suchas binding assays, fluorescence polarization assays, FRET-basedcoactivator recruitment assays (see, generally, Glickman et al., J.Biomolecular Screening (2002), Vol. 7, No. 1, pp. 3-10), as well as cellbased assays including the co-transfection assay, the use of LBD-Gal 4chimeras and protein-protein interaction assays, (see, Lehmann. et al.,J. Biol Chem. (1997), Vol. 272, No. 6, pp. 3137-3140).

Compounds of the present invention show unexpected advantages overcompounds previously disclosed in the art, such as those disclosed inPCT Publ. No. WO 2007/002563. The present compounds have been shown inan assay(s), such as those described below, to have a desirable partialLXR agonist character with increased potency in human whole blood andlow LXRa efficacy. Additionally, the compounds of the present inventionalso exhibit metabolic stability in a human liver microsomal assay. Suchcompounds should be more useful in the treatment, inhibition oramelioration of one or more diseases or disorders that are discussedherein.

Example A Scintillation Proximity Assay (SPA)

The SPA assay measures the radioactive signal generated by the bindingof ³H-24,25-epoxycholesterol to LXRα-RXR_(α) or LXR_(β)-RXR_(α)heterodimers. The basis of the assay is the use of SPA beads containinga scintillant, such that when binding to the receptor brings the labeledligand into proximity with the bead, the energy from the labelstimulates the scintillant to emit light. The light is measured using astandard microplate scintillation reader. The ability of a ligand tobind to a receptor can be measured by assessing the degree to which thecompound can compete off a radiolabelled ligand with known affinity forthe receptor.

Required Materials:

-   -   1. Label: ³H-24,25-epoxy-cholesterol (NEN Life Science        Products/Perkin Elemer))    -   2. LXR_(α) lysate: Baculovirus expressed LXR_(α)/RXR heterodimer        both with a 6-HIS tag produced as a crude lysate    -   3. LXRβ lysate: Baculovirus expressed LXR_(β)/RXR heterodimer        both with a 6-HIS tag produced as a crude lysate    -   4. SPA beads: Ysi copper His-tag SPA beads (Amersham)    -   5. Plates: Non-binding surface 384-well plate (Corning)    -   6. Protein lysate dilution buffer: (20 mM Tris-HCl pH 7.9, 500        mM NaCl, 5 mM Imidazole).    -   7. 2×SPA Buffer: (40 mM K₂HPO₄/KH₂PO₄ pH7.3, 100 mM NaCl, 0.05%        Tween 20, 20% Glycerol, 4 mM EDTA)    -   8. 2×SPA Buffer w/o EDTA: (40 mM K₂HPO₄/KH₂PO₄ pH7.3, 100 mM        NaCl, 0.05% Tween 20, 20% Glycerol)

Stock Solutions

-   -   0.5 M K₂HPO₄/KH₂PO₄ pH 7.3    -   0.5 M EDTA pH 8.0    -   5 M NaCl    -   10% Tween-20    -   Glycerol

Preparation of Protein Lysates

Baculovirus expression plasmids for human RXR α □□□ (accession NoNM_002957), LXR_(α) (accession No U22662), and LXR_(β) (accession NoU07132) were made by cloning the appropriate full-length cDNAs into thepBacPakhis2 vector (Clontech, CA) following standard procedures.Insertion of the cDNAs into the pBAcPakhis2 vector polylinker created anin frame fusion to the cDNA to an N-terminal poly-His tag present inpBacPakhis1. Correct cloning was confirmed by restriction mapping,and/or sequencing.

Cell lysates were prepared by infecting healthy, Sf9 insect cells at adensity of approximately 1.25×10⁶/ml at 27° C., in a total volume of 500mL per 1 L sized spinner flasks, cultured under standard conditions. Toprepare the LXR_(α) lysate, insect cells were co-transfected with theLXR_(α) expression cassette at an M.O.I. of 2.0 and with the RXRexpression cassette at a M.O.I. of approximately 1.0. To prepare theLXR_(β) lysate, insect cells were co-transfected with the LXR_(β)expression cassette at an M.O.I of approximately 2.0 and with the RXRexpression cassette at a M.O.I. of approximately 1.0. In both casescells were incubated for 48 hours at 27° C. with constant shaking priorto harvesting.

After incubation, cells were harvested by centrifugation and pelleted.Cell pellets were resuspended in two volumes of ice-cold freshlyprepared extraction buffer (20 mM Tris pH 8.0, 10 mM Imidazole, 400 mMNaCl, containing one EDTA free protease inhibitor tablet (Roche CatalogNo: 1836170) per 10 ml of extraction buffer).

Cells were homogenized slowly on ice using a Dounce homogenizer toachieve 80-90% cell lysis. The homogenate was centrifuged in apre-chilled rotor (Ti50 or Ti70, or equivalent) at 45,000 rpm for 30minutes at 4° C. Aliquots of the supernatant were frozen on dry ice andstored frozen at −80° C. until quantification and quality control.Aliquots of the lysates were tested in the SPA assay to ensure lot tolot consistency, and via SDS-PAGE analysis after purification usingNi-NTA Resin (Qiagen) and adjusted for protein concentration andexpression level prior to use in screening assays.

Preparation of Screening Reagents

[³H] 24,25 Epoxycholesterol (EC) solution: For a single 384-well plate(or 400 wells), 21 μL of [³H] EC (specific activity 76.5 Ci/mmol,concentration 3.2 mCi/mL) was added to 4.4 mL of 2×SPA buffer to providefor a final concentration of 200 nM. For each additional 384-well plate,an additional 19.1 μL of [³H] EC was added to 4.0 mL of additional 2×SPAbuffer. The final concentration of [³H] EC in the well was 50 nM.

LXR_(α) lysate (prepared as above) was diluted with protein lysatedilution buffer. 1400 μL of diluted LXR_(α) lysate was prepared per384-well plate, (or 200 wells) and 1120 μL of diluted LXR_(α) lysate wasprepared for each additional 384-well plate.

LXRβ lysate (prepared as above) was diluted with protein lysate dilutionbuffer. 1400 μL of diluted LXR_(β) lysate was prepared per 384-wellplate, (or 200 wells) and 1120 μL of diluted LXRβ lysate was preparedfor each additional 384-well plate.

SPA bead solution: For a 384-well plate (or 400 wells), 3.75 mL of 2×SPAbuffer w/o EDTA, 2.25 mL of H₂O, and 1.5 mL of Ysi His-tag SPA beads(vortex well before taking) were mixed together. For each additional384-well plate, an additional 3.5 mL of 2×SPA buffer w/o EDTA, 2.1 mL ofH₂O, and 1.4 mL of Ysi His-tag SPA beads were mixed together.

Procedure:

Appropriate dilutions of each compound were prepared in a 96-well plateand pipetted into the appropriate wells of a 384 well plate at 3.5 μlper well.

-   -   9.1 μL of [³H] EC was added to each well of column 2-23 of the        multiwell plate.    -   5 μl of diluted LXR_(α) lysate was added to each well of column        2-23 on odd rows of the multiwell plate.

5 μL of diluted LXR_(β) lysate was added to each well of column 2-23 oneven rows of the multiwell plate.

17.5 μL of SPA bead solution was added to each well of column 2-23 ofthe multiwell plate.

The plates were covered with clear sealer and placed in an incubator atambient temperature for approximately 30 minutes.

After incubation plates were analyzed using a luminescent plate reader(MicroBeta, Wallac) using the program n ABASE 3H_384DPM. The setting forn ABASE 3H_384 DPM was:

-   -   Counting Mode: DPM;    -   Sample Type: SPA;    -   ParaLux Mode: low background;    -   Count time: 30 sec.

Assays for LXR_(α) and LXR_(β) were performed in the identical manner.The determined Ki represents the average of at least two independentdose response experiments. The binding affinity for each compound may bedetermined by non-linear regression analysis using the one sitecompetition formula to determine the IC₅₀ where:

Y=Bottom+(Top−Bottom)/(1+10^(X-logIC50)).

The Ki is than calculated using the Cheng and Prusoff equation where:

Ki=IC₅₀/(1+[Concentration of Ligand]/Kd of Ligand).

For this assay, typically the Concentration of Ligand=50 nM and the Kdof EC for the receptor is 200 nM as determined by saturation binding.

The compounds of the invention demonstrated the ability to bind toLXR_(α) and/or LXR_(β), when tested in this assay.

Example B Co-Transfection Assay

To measure the ability of compounds to activate or inhibit thetranscriptional activity of LXR in a cell based assay, theco-transfection assay was used. It has been shown that LXR functions asa heterodimer with RXR. For the co-transfection assay, expressionplasmids for LXRα and LXRβ are introduced separately via transienttransfection into mammalian cells along with a luciferase reporterplasmid that contains one copy of a DNA sequence that is bound byLXR-RXR heterodimers (LXRE; Willy, P. et. al. 1995). LXRs heterodimerizewith the endogenous RXR. Treatment of transfected cells with an LXRagonist increases the transcriptional activity of LXR, which is measuredby an increase in luciferase activity. Similarly, LXR antagonistactivity can be measured by determining the ability of a compound tocompetitively inhibit the activity of a LXR agonist.

Required Materials

CV-1 African Green Monkey Kidney Cells

Co-transfection expression plasmids, comprising full-length LXR_(α)(pCMX-h LXR_(α)□ or LXR_(β) (pCMX-hLXR_(β)), reporter plasmid(LXREx1-Tk-Luciferase), and control (pCMX-Galactosidase expressionvector) (Willey et al. Genes & Development 9 1033-1045 (1995)).

Transfection reagent such as FuGENE6 (Roche).

-   -   1× cell lysis buffer:        -   22.4 mM Tricine pH 8.0        -   0.56 mM EGTA pH 8.0        -   5.6 mM MgSO₄        -   0.6% Triton X-100        -   5.6% glycerol    -   10× luciferase substrate solution:        -   10 mM HEPES pH 6.5        -   2.75 mM D-Luciferin        -   0.75 mM Coenzyme-A        -   3.7 mM ATP        -   96 mM DTT

Preparation of Screening Reagents

CV-1 cells were prepared 24 hours prior to the experiment by platingthem into T-175 flasks or 500 cm² dishes in order to achieve 70-80%confluency on the day of the transfection. The number of cells to betransfected was determined by the number of plates to be screened. Eachwell of a 384 well plate requires 8000 cells. DNA Transfection Reagentwas prepared by mixing the required plasmid DNAs with a cationic lipidtransfection reagent FuGENE6 (Roche) by following the instructionsprovided with the reagents. Optimal DNA amounts were determinedempirically per cell line and size of vessel to be transfected. For eachT175 cm² flask a total of 20 μg of DNA, 60 μl of Fugene 6 and 1 ml ofOptimem was mixed and added. Cells were then incubated at least 5 hoursat 37° C. to prepare screening cells.

Luciferase assay reagent was prepared by combining before use:

-   -   1 part of 10× Luciferase substrate solution    -   9 parts of 1× cell lysis buffer.

Procedure

Assay plates were prepared by dispensing 5 μL of compound per well of a384 well plate to achieve final compound concentration of 10 μM and nomore than 0.5% DMSO. Media was removed from the screening cells, thecells trypsinized, harvested cells by centrifugation, counted, andplated at a density of approximately 8000 cells per well in the 384 wellassay plate prepared above in a volume of about 45 μL. Assay platescontaining both compounds and screening cells (50 μL in total volume)were incubated for 20 hours at 37° C.

After incubation with compounds, media was removed from the cells andluciferase assay reagent (30 μL/well) added. After ˜2 minutes at ambienttemperature, the assay plates were read on a luminometer (PE BiosystemsNorthstar reader with on-board injectors, or equivalent).

The LXR/LXRE co-transfection assay can be used to establish theEC₅₀/IC₅₀ values for potency and percent activity or inhibition forefficacy. Efficacy defines the activity of a compound relative to a highcontrol((N-(3-(4-fluorophenyl)-(naphthalene-2-sulfonyl)amino)propyl)-2,2-dimethylpropionamide))or a low control (DMSO/vehicle). The dose response curves are generatedfrom a 10 point curve with concentrations differing by ½ LOG units. Eachpoint represents the average of 4 wells of data from a 384 well plate.

The data from this assay is fitted to the following equation, from whichthe EC₅₀ value may be solved:

Y=Bottom+(Top−Bottom)/(1+10^(((logEC50-X)*HillSlope))).

The EC₅₀/IC₅₀ is therefore defined as the concentration at which anagonist or antagonist elicits a response that is half way between theTop (maximum) and Bottom (baseline) values. The EC₅₀/IC₅₀ valuesrepresented are the averages of at least 2 and normally 3 independentexperiments. The determination of the relative efficacy or % control foran agonist is by comparison to the maximum response achieved by((N-(3-((4-fluorophenyl)-(naphthalene-2-sulfonyl)-amino)propyl)-2,2-dimethylpropionamide)that is measured individually in each dose response experiment.

For the antagonist assay, a LXR agonist can be added to each well of a384 well plate to elicit a response. The % inhibition for eachantagonist is therefore a measurement of the inhibition of the activityof the agonist. In this example, 100% inhibition would indicate that theactivity of a specific concentration of LXR agonist has been reduced tobaseline levels, defined as the activity of the assay in the presence ofDMSO only.

Compounds of the present invention were tested in the LXRα assaydescribed immediately above and were shown to have efficacy of less thanor equal to 25% of the control compound.

Compounds of the present invention were tested in the LXRIβ assaydescribed immediately above and were shown to have efficacy of greaterthan or equal to 30% of the control compound.

Example C Human Whole Blood Assay

Human whole blood was collected in EDTA containing tubes and 0.5 mLaliquots were immediately mixed in a 96 well block with the appropriateserial dilution of test compound, in 0.5% DMSO. Compounds were incubatedwith blood at 37° C. with constant rocking for 4 hours. Afterincubation, cells were lysed in ABI Nucleic Acid Purification LysisSolution (Applied Biosystems catalog #4305895) and frozen at −80° C.Following cell lysis, total RNA was purified using an ABI 6100 RNA prepstation according to the protocol provided by the manufacturer. cDNAswere synthesized, and mRNAs were quantitated using SYBR-GreenQuantitative PCR (Q-PCR) on an ABI Prism 7900HT Sequence DetectionSystem and reagents from Quanta Bioscience Inc (Quanta Biosciencecatalog #95047 and 95055).

TABLE 2.1 Primers Used for mRNA Quantitation by RT-PCR GeneForward Primer Reverse Primer ABCA1 GGTGATGTTTCTGA TGTCCTCATACCAGCCAATGTGA TTGAGAGAC (SEQ ID No. 1) (SEQ ID No. 2) ABCG1 GACTGCGTGTCCTGGATGGGGGCATGAT CAAAATC GACAATG (SEQ ID No. 3) (SEQ ID No. 4) L-30GCTGGAGTCGATCA CCAATTTCGCTTTG ACTCTAGG CCTTGTC (SEQ ID No. 5)(SEQ ID No. 6) B2M GGCTATCCAGCGTA CGGCAGGCATACTC CTCCAAA ATCTTTTT(SEQ ID No. 7) (SEQ ID No. 8)

The quantity of each mRNA was determined by the ΔΔCT method (Michael W.Pfaffl. A new mathematical model for relative quantification inreal-time RT-PCR. Nucleic Acids Research, 2001, Vol. 29, No. 9 e45) andnormalized to the quantity of two control mRNAs, i.e. ribosomal proteinL-30 (L-30) and beta-2-microglobulin (B2M). The induction of ABCA1 andABCG1 by test compound was graphed as a percent of the referencecompound,2-(4-(5-(5-cyano-1-(2,4-difluorobenzyl)-6-oxo-4-(trifluoromethyl)-1,6-dihydropyridin-2-yl)thiophen-2-yl)-3-methylphenyl)aceticacid, and potency (EC₅₀) and activity (% MAX) were calculated by fittinga sigmoidal response curve as a function of log transformed compoundconcentration using XLFit software according to the equationy=A+((B−A)/(1+((C/x)̂D))). The full LXRα and LXRIβ pan agonist2-(4-(5-(5-cyano-1-(2,4-difluorobenzyl)-6-oxo-4-(trifluoromethyl)-1,6-dihydropyridin-2-yl)thiophen-2-yl)-3-methylphenyl)aceticacid, (EC₅₀ 1-2 μM) was used as a reference compound and its maximalactivity was defined as 100%.

Compounds of the present invention were tested in the assay describedimmediately above and were shown to have a potency generally less than1,000 nM, preferably, less than 100 nM, more preferably less than 20 nM.

Example D High Throughput (HT) Metabolic Stability in Human Microsomes

The metabolic stability assay evaluates CYP-mediated metabolic stabilityin vitro using Human liver microsomes after a ten-minute incubation.

Test compound is received as a 3.5 mM stock solution in 100 percentDMSO. Compound is diluted to create a 50 μM acetonitrile (ACN) solutioncontaining 1.4% DMSO, which is then used as a 100× stock for incubationwith human liver microsomes. Each compound is tested in duplicate.Compound, NADPH and liver microsome solutions are combined forincubation in three steps:

-   -   1) 152 μl of liver microsome suspension, protein concentration        of 1.1 mg/ml in 100 mM NaP_(i), pH 7.4, 5 mM MgCl₂ buffer, is        pre-warmed at 37° C.    -   2) 1.7 μl of 50 μM compound (98.6% ACN, 1.4% DMSO) is added to        the same tube and pre-incubated at 37° C. for 5 minutes.    -   3) The reaction is initiated by the addition of 17 μl of        pre-warmed 10 mM NADPH solution in 100 mM NaP_(i), pH 7.4.

Reaction components are mixed well, and 75 μl are immediatelytransferred into 150 μl quench/stop solution (zero-time point, T₀).Reactions are incubated at 37° C. for 10 minutes and then an additional75 μl aliquot is transferred into 150 μl quench solution. Acetonitrilecontaining 100 μM DMN (a UV standard for injection quality control), isused as the quench solution to terminate metabolic reactions.

Quenched mixtures are centrifuged at 1500 rpm (˜500×g) in an AllegraX-12 centrifuge, SX4750 rotor (Beckman Coulter Inc., Fullerton, Calif.)for fifteen minutes to pellet denatured microsomes. A volume of 90 μl ofsupernatant extract, containing the mixture of parent compound and itsmetabolites, is then transferred to a separate 96-well plate forUV-LC/MS-MS analysis to determine the percent of parent compound that isremaining in the mixture.

UV-LC/MS-MS Sample Analysis—Structural Integrity Pre-Analysis

The Metabolic Stability structural integrity pre-analysis is used toassess the purity of compounds being assayed. Compounds are received in96-well plates as 57 μl of a 3.5 mM DMSO solution. The 3.5 mM compoundDMSO stock solutions are diluted 18-fold with a solution containingequal volumes of acetonitrile, isopropanol, and MilliQ-H₂O. Theresulting solutions (200 μM) are analyzed for structural integrity byLC-UV/MS on a Thermo LCQ Deca XP Plus ion trap mass spectrometer, usinga Waters XBridge C18, 5 μm, 2×50 mm column with a Waters Sentry 2.1 mmguard column, and the LC conditions described in the table below, with a5 μl injection and a flow rate of 1 ml/min. The acquired data reflectpurity by UV absorbance at 220 nm. Only results for those compounds withpurity greater than 50% are reported.

Metabolic Stability - Structural Integrity HPLC Gradient* Gradient Time(min) A % B % 0.00 100 0 4.00 0 100 5.00 0 100 5.10 100 0 6.00 100 0*Mobile Phase for structural integrity pre-analysis: (A) 98% water, 2%acetonitrile with 10 mM ammonium acetate; (B) 10% water, 90%acetonitrile with 10 mM ammonium acetate

Sample Analysis—Incubated Samples

MS/MS condition optimization is conducted on a Thermo TSQ Quantumtriple-quadropole mass spectrometer equipped with a Heated-electrospray(H-ESI) source by automated infusion to obtain the SRM transitions andtheir corresponding collision energy values. Compound solutions at aconcentration of 20 μM in 1:1 methanol:water are infused at a flow rateof 90 μL/min, then combined with the mobile phase at a flow rate of 50μL/min before being introduced into the source. All compounds areoptimized first using mobile phase A and B (50% A and 50% B), and ifnecessary, using mobile phase C and D (also with a 50:50 composition).The optimized parameters, including polarity, SRM transition andcollision energy, are stored in a Microsoft Access database.

The mass spectrometric conditions obtained from automated infusion areused to analyze incubation samples from the Metabolic Stability assay.The injection volume is 5 μl and the flow rate is 0.8 ml/min. Thegradient used is show in the table below. All samples are injected withthe gradient using mobile phase A and B first. If necessary (forinstance, for chromatographic reasons), samples are re-injected with thesame gradient, but using mobile phase C and D. All LC-MS/MS analysisparameters are captured electronically in the raw data files.

Metabolic Stability - Sample Analysis Gradient* A % B % Gradient Time(min) (or C %) (or D %) 0.00 95 5 0.20 95 5 0.30 0 100 1.05 0 100 1.1095 5 1.50 95 5 *Mobile Phase for reaction sample analysis: (A) 98%water, 2% acetonitrile with 0.1% formic acid; (B) 2% water, 98%acetonitrile with 0.1% formic acid; (C) 0.1% ammonium hydroxide inwater; (D) 0.1% ammonium hydroxide in acetonitrile

Peak integration is performed with the Xcalibur™ software. The percentremaining calculation is performed by comparing the LC-MS/MS peak areasfrom the T_(10minute) samples to those from the T_(10minute) samples foreach compound.

Compounds of the present invention were tested in the assay describedimmediately above and were shown to have greater than 80% of parentcompound remaining at 10 minutes.

The following Table 1 presents the results of the LXR/LXREco-transfection assay that measures LXRα EC50 and efficacy, the humanwhole blood assay (hWBA) that measures the ability of the compounds tobind to LXR and induce ABCA1 gene expression relative to a referencecompound, and the microsome metabolic stability assay. EC₅₀ values aregiven in ranges where A is ≦100 nM, B is 101 nM to <1,000 nM and Cis >1,000 nM.

TABLE 1 hWBA hWBA LXRα LXRα hABCA1 hABCA1 Human EC50 Eff L30 L30Microsome % Structure # (nm) (%) EC50 (nM) Maz P (%) Remaining

3 A 6 A 16 >80%

11 C — A 16 >80%

21 A 18 A 17 >80%

2 A 10 A 18 >80%

17 B 13 A 20 >80%

8 A 10 A 23 >80%

14 A 15 A 24 >80%

1 B 13 A 28 >80%

9 A 20 A 28 >80%

12 A 13 A 32 >80%

7 A 15 A 33 >80%

4 A 12 A 34 >80%

20 B 15 A 35 >80%

19 C — A 35 >80%

16 A 19 A 41 >80%

5 A 25 A 43 >80%

6 A 25 A 35 >80%

15 A 8 A 47 >80%

13 A 18 A 50 >80%

18 A 17 A 51 >80%

Compound No. 19, Table 1, WO 2007/002563 B 38 C 55 >80%

The above representative data illustrates the unexpected desirablepartial LXR agonist character, increased potency in human whole blood,low LXRa efficacy and metabolic stability in a human liver microsomalassay of the compounds of the present invention in comparison tocompounds previously disclosed in the art, such as those disclosed inPCT Publ. No. WO 2007/002563.

Example E In Vivo Potency and Maximum ABCG1 Induction in CynomolgusMonkey

Compounds of the present invention were tested for their ability toinduce the mRNA for the LXR target gene ABCG1 in blood cells whenadministered orally to cynomolgus monkeys.

Test compounds were formulated in 0.5% carboxymethyl cellulose (CMC,Sigma) and 2% Tween 80 (Sigma) by trituration. Each treatment groupcontained three male monkeys, each weighing 3.0-6.0 kg at the start ofthe study. Test compounds were formulated fresh each morning in vehicle,and animals that had been fasted the previous night were dosed between 7and 7:30 AM by po gavage. For baseline blood cell mRNA determinations, 1ml of venous blood was collected in an EDTA dry-coated tube and 1 volumeof Dulbecco's Phosphate Buffered Saline without calcium or magnesium and2 volumes of Nucleic Acid Purification Lysis Solution (AppliedBiosystems, Inc.) were added. Samples were frozen at −80° C. prior toRNA isolation and analysis. Test compounds were dosed following thebaseline sample collection. At 5 hr post-dose, venous blood wascollected and processed as above for RNA determinations. An additional0.5 ml of blood was also collected and analyzed for compound plasmaconcentrations.

RNA Isolation.

Frozen samples were allowed to thaw at room temperature, and then placedon ice. Total RNA was isolated on the ABI 6100 using a pre-loadedprotocol according to the manufacturer's instructions (ABI Manual#4332809 Rev. B).

cDNA Synthesis and Q-PCR Reactions.

qScript cDNA Synthesis Kit from Quanta Biosciences, Inc. was used togenerate first-strand cDNA from each total RNA sample. 20 μl Reactionswere carried out in 96-well Eppendorf AG twin-tec PCR plates on a MJResearch, Inc. model PTC-200 DNA Engine. Approximately 500 ng of totalRNA was used for each reaction. Reactions were composed as follows: 4 μl5× q-Script Reaction Mix plus 3-5 μl nuclease-free water plus 1 μlq-Script Reverse Transcriptase plus 10-12 μl input RNA. After mixing,reactions were centrifuged at 3750 rpm for 2 minutes at room temperatureand run on a “q-Script” protocol (25° C. for 5 minutes, followed by 42°C. for 30 minutes, and then 85° C. for 5 minutes) in a MJ Researchthermocycler. Each reaction was diluted with 30 μl of nuclease-freewater and used immediately for SYBR-green Q-PCR or stored at −20° C.SYBR-Green Q-PCR reactions were performed as follows. Reaction mixturesfor the LXR target gene, ABCG1, and the internal standard normalizationgene ribosomal protein L30 were prepared using validated forward/reverseprimer sets [ABCG1 (X-Mmul-ABCG1-F1 and X-Mmul-ABCG1-R1); ribosomalprotein L30 (SYBR-hL30-F1 and SYBR-hL30-R1)]. Information on sequencesof these primer sets were obtained from the “Primer Bank” Web Site(http://pga.mgh.harvard.edu/primerbank/index.html) for L30 (Primer Bank#4506631a1), and from Exelixis, Inc. (South San Francisco, Calif., USA)for ABCG1 (Mmul_ABCG1.TaqMan F1/R1).

Primer Sequences

Mmul_ABCG1.TaqMan.F1 (SEQ ID No. 9) 5′-ACGCAGACAGCACTGGTGAA-3′Mmul_ABCG1.TaqMan.R1 (SEQ ID No. 10) 5′-CTTCCCTCCACCTGGAACCT-3′ hL30-F1(SEQ ID No. 5) 5′-GCTGGAGTCGATCAACTCTAGG-3′ hL30-R2 (SEQ ID No. 6)5′-CCAATTTCGCTTTGCCTTGTC-3′

SYBR-Green reactions were assembled as follows. Per reaction, 10 μl 2×PowerSYBR® Green Supermix (Applied Biosystems Catalog #4367659) plus 2μl of 10 μM Gene Specific Forward/Reverse Primer Mix (finalconcentration of primers is 500 nM each) plus 4 μl water were mixedtogether with 4 μl of diluted RT reaction. Reaction mixes werecentrifuged at 3750 rpm for 2 minutes at room temperature and run on anApplied Biosystems model 7900HT SDS-Taqman System.

Calculation of Relative mRNA Quantities, and In Vivo Compound Potenciesand Maximal Activites.

The relative amount of ABCG1 mRNA was calculated using the secondderivative comparative Ct method (2^(−ΔΔ)Ct). Quantification wasobtained after normalization to ribosomal protein L30 mRNA. Each samplewas tested in duplicate and the average Ct was used for calculations.

Calculation of In Vivo Potency of Compounds in Cynomolgus Monkeys.

The concentration of test compound in plasma from each individual animalat 5 hrs post-dose was plotted vs. the fold induction vs. baseline ofABCG1 mRNA in blood cells for each animal at 5 hrs post-dose for alldose groups. Data from all dose groups were included on the same plotfor each compound. The in vivo potency (EC₅₀) and maximum induction ofABCG1 mRNA for each compound was determined by non-linear curve fittingusing a sigmoidal dose-response equation (Graphpad Prism 4.03 software).

Quantitation of Plasma Compound Concentrations by LC/MS/MS.

The following are details of the liquid chromatography with tandem massspectrometry (LC/MS/MS)-based bioanalytical methods used to quantitatetest compounds in cynomolgus monkey plasma.

The LC/MS/MS analysis of the test compound was conducted against astandard curve ranging from 1 to 5000 nM. The standard curves werefitted with a linear regression weighted by reciprocal squared (1/x²).Standards were analyzed in single replicates. Quality control sampleswere prepared in blank biological matrix at 3 concentrations within therange of the standard curve and were analyzed as replicates within eachanalytical set. The determined concentrations of more than 75% of theQCs were within 20% of their nominal values.

Sample preparation was conducted on Janus 8-tip and Janus Mini 96-tipautomated liquid handlers. Aliquots (50 μL) of the biological matrix(plasma) from in vivo studies and standard/QC samples were treated with1 M ammonium carbonate in water pH 9.2 unadjusted (50 μL) containing 200nM of two internal standards (IS), followed by 300 μL methyl t-butylether (MTBE) and partitioned by liquid-liquid extraction (LLE) usingforty fill-expel tip repetitions for approximately 3 minutes. Theaqueous and organic layers were then centrifuged for 2-min at 3900 rpm.The top organic layer extraction solvent MTBE (250 μL) was removed toanother clean 96-well plate and placed in a nitrogen evaporator for 15min at 40° C. to dryness. Aliquots (100 μL) were used to reconstitutethe dry extracts with mobile phase consisting of 1:1 acetonitrile/water.A 10 μL aliquot was first injected onto the high-Turboflow performanceliquid chromatography (HTLC) extraction column, then eluted onto asecond high-performance liquid chromatography (HPLC) column forLC/MS/MS-based analysis.

The LC system used for all analyses was an Aria TX-2 (Thermo Scientific,Waltham, Mass., USA) HPLC system consisting of 8 Shimadzu LC10AD pumpswith 2 SCL-10AVP System Controllers (Columbia, Md., USA) and a dual armCTC Analytics HTS PAL autosampler (Switzerland) equipped with a coolingstack that maintained samples at 10° C. during analysis. The HPLCon-line extraction column was a Cyclone-P mixed polymer (0.5×50 mm, 50particle, Thermo Scientific, Waltham, Mass., USA) kept at roomtemperature. The HPLC C18 analytical column used was an XBridge C18 (2.1mm×50 mm, 5 μM particle, Waters Corporation, Milford, Mass., USA) keptat room temperature. The mobile phase, which consisted of 0.1% formicacid in water (A) and 0.1% formic acid in acetonitrile (B), wasdelivered at a flow rate of 1.5 mL/min to the HTLC on-line extractioncolumn and at 0.5 mL/min to the HPLC C18 analytical column. These flowrates change during the transfer step between 0.5 to 1.0 min. Theretention times for the test compounds and internal standards wererecorded. The total analysis time was 5.0 min. The gradients aresummarized in the following tables.

Mobile Phase Gradient for HPLC C18 XBridge Analytical Column Flow RateTime (min) % A % B (mL/min) Curve 0 (Initial) 95 5 0.5 isocratic 0.50 955 0.3 isocratic 1.00 95 5 0.3 isocratic 1.10 95 5 0.5 isocratic 2.00 595 0.5 linear 2.50 5 95 0.5 isocratic 2.60 95 5 0.5 step

Mobile Phase Gradient for HTLC On-Line Extraction Cyclone-P Column FlowRate Time (min) % A % B (mL/min) Curve 0 (Initial) 100 0 1.5 isocratic

Mobile Phase Gradient for HPLC C18 XBridge Analytical Column Flow RateTime (min) % A % B (mL/min) Curve 0.50 100 0 0.2 isocratic 1.00 100 00.2 isocratic 1.10 0 100 1.5 step 2.00 0 100 1.5 isocratic 2.10 50 501.5 step 2.50 50 50 1.5 isocratic 2.60 100 0 1.5 step

The mass spectrometer used for all analyses was a Finnigan Quantum Ultratandem mass spectrometer (Thermo Scientific, Waltham, Mass., USA)equipped with a heated electrospray interface operating in both positiveand negative ionization modes. Ultra-high-purity (UHP) nitrogen was usedas the sheath and aux gases at flow rates of 55 psi for sheath and 25units for aux. The desolvation temperature was 350° C. and the sourcetemperature was 350° C. Detection of each analyte was achieved throughselected reaction monitoring (SRM). Positive ionization mode was used toquantitate the test compound and the internal standard. UHP argon at apressure of 1.5×10⁻³ torr was maintained in the collision cell ofquadrupole 2. The transitions monitored for a compound of the presentinvention and and its internal standard were recorded.

The in vivo potency in cynomologus monkey of test compounds, and themaximum ABCG1 induction derived from plots of plasma compoundconcentration at 5 hrs post-dose vs. fold mRNA induction at 5 hrspost-dose are shown in Table 2 below.

TABLE 2 Maximum ABCG1 Induction Compound EC₅₀ ± S.E. (nM) (fold vs.baseline ± S.E.) Compound No. 19, 630 ± 168 12.4 ± 1.8  Table 1, WO2007/002563 Example 4 141 ± 53  5.9 ± 0.8 Present Invention Example 9 11± 6 8.9 ± 0.9 Present Invention

Examples 4 and 9 of the present invention are about four and sixty foldmore potent than a compound know in the art after dosing in cynomologusmonkeys. Similarly, Examples 4 and 9 induce ABCG1 mRNA in blood to amaximum of about six and nine fold compared to about twelve fold for acompound known in the art thereby demonstrating partial LXR activity incynomologus monkey blood.

It is understood that the examples and embodiments described herein arefor illustrative purposes only and that various modifications or changesin light thereof will be suggested to persons skilled in the art and areto be incorporated within the spirit and purview of this application andscope of the appended claims. All publications, patents, and patentapplications cited herein are hereby incorporated herein by referencefor all purposes.

We claim:
 1. The compound2-(1-(3′-fluoro-4′-(hydroxymethyl)-5′-(methylsulfonyl)biphenyl-4-yl)-2-(2-(2-fluorophenyl)propan-2-yl)-1H-imidazol-4-yl)propan-2-olof formula:

or an isotopically-labeled compound, or a pharmaceutically acceptablesalt thereof.
 2. A pharmaceutical composition comprising the compound ofclaim 1, or an isotopically-labeled compound, or a pharmaceuticallyacceptable salt thereof, and one or more pharmaceutically acceptablecarriers.
 3. A method of treating a disease or disorder comprisingadministering to a subject in need thereof a therapeutically effectiveamount of the compound of claim 1, or an isotopically-labeled compound,or a pharmaceutically acceptable salt thereof, wherein the disease ordisorder is insulin resistance, osteoarthritis, stroke, hyperglycemia,psoriasis, age and UV exposure-related skin wrinkling, diabetes, cancer,Alzheimer's disease, inflammation, immunological disorders, lipiddisorders, obesity, macular degeneration, conditions characterized by aperturbed epidermal barrier function, conditions of disturbeddifferentiation or excess proliferation of the epidermis or mucousmembrane, or cardiovascular disorders.
 4. The method of claim 3, whereinthe disease or disorder is a cardiovascular disorder, a lipid disorder,diabetes, or Alzheimer's disease.
 5. The method of claim 4, wherein thedisease or disorder is a cardiovascular disorder.
 6. The method of claim5, wherein said cardiovascular disorder is atherosclerosis.
 7. Themethod of claim 4, wherein the disease or disorder is a lipid disorder.8. The method of claim 7, wherein said lipid disorder is dyslipidemia.9. The method of claim 3 wherein the disease or disorder is diabetes.10. The method of claim 3 wherein the disease or disorder is Alzheimer'sdisease.